Compounds for inhibiting insulin secretion and methods related thereto

ABSTRACT

Compounds, compositions and methods for altering insulin secretion, particularly in the context of treatment of subjects having, or suspected of being at risk for having, diabetes mellitus. The compounds have the following struture (I):  
                 
 
including stereoisomers, prodrugs and pharmaceutically acceptable salts thereof, wherein W 1 , W 2 , X, R 1 , R 2 , R 3 , R 4 , m and n are defined herein.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication No. 60/355,389 filed Feb. 8, 2002, where this provisionalapplication is incorporated herein by reference in its entirety.

TECHNICAL FIELD

The invention generally relates to compounds that alter insulinsecretion, as well as to composition and methods related thereto.

BACKGROUND OF THE INVENTION

Type 2 diabetes mellitus, or “late onset” diabetes, is a common,degenerative disease affecting 5 to 10 percent of the population indeveloped countries. The propensity for developing type 2 diabetesmellitus (“type 2 DM”) is reportedly maternally inherited, suggesting amitochondrial genetic involvement. (Alcolado, J. C. and Alcolado, R.,Br. Med. J. 302:1178-1180 (1991); Reny, S. L., International J. Epidem.23:886-890 (1994)). Diabetes is a heterogeneous disorder with a stronggenetic component; monozygotic twins are highly concordant and there isa high incidence of the disease among first degree relatives of affectedindividuals.

At the cellular level, the pathologic phenotype that may becharacteristic of the presence of, or risk for predisposition to, lateonset diabetes mellitus includes the presence of one or more indicatorsof altered mitochondrial respiratory function, for example impairedinsulin secretion, decreased ATP synthesis and increased levels ofreactive oxygen species. Studies have shown that type 2 DM may bepreceded by or associated with certain related disorders. For example,it is estimated that forty million individuals in the U.S. suffer fromimpaired glucose tolerance (IGT). Following a glucose load, circulatingglucose concentrations in IGT patients rise to higher levels, and returnto baseline levels more slowly, than in unaffected individuals. A smallpercentage of IGT individuals (5-10%) progress to non-insulin dependentdiabetes (NIDDM) each year. This form of diabetes mellitus, type 2 DM,is associated with decreased release of insulin by pancreatic beta cellsand a decreased end-organ response to insulin. Other symptoms ofdiabetes mellitus and conditions that precede or are associated withdiabetes mellitus include obesity, vascular pathologies, peripheral andsensory neuropathies and blindness.

Glucose-mediated insulin secretion from the pancreatic beta cell istriggered by a complex sequence of intracellular events. Glucose istaken up by the beta cell via GLUT-2 glucose transporters; it issubsequently phosphorylated by glucokinase to glucose-6-phosphate, whichenters the glycolytic pathway. The reducing equivalents (NADH) andsubstrate (pyruvate) produced through glycolysis enter the mitochondriaand fuel increased respiration and oxidative phosphorylation. Theconsequent rise in cellular ATP levels triggers closure of the K⁺-ATPchannels at the plasma membrane, depolarizing the membrane andpermitting influx of calcium. Calcium appears to have two main roles:stimulating release of insulin from the cells (e.g., Kennedy et al.,1996 J. Clin. Invest. 98:2524; Maechler et al., 1997 EMBO J. 16:3833),and acting as a “feed-forward” regulator of mitochondrial ATP production(e.g., Cox and Matlib, 1993 Trends Pharmacol. Sci. 14:408). The latteris accomplished by mitochondrial uptake of calcium through themitochondrial calcium uniporter (e.g., Newgard et al., 1995 Ann. Rev.Biochem. 64:689; Magnus et al., 1998 Am. J. Physiol. 274:C1174-C1184).The rise in mitochondrial calcium stimulates respiration and oxidativephosphorylation through stimulation of calcium-sensitive dehydrogenase(Rutter et al., 1988 Biochem. J. 252:181; Rutter et al., 1993 J. Biol.Chem. 268:22385). However, the rise in mitochondrial calcium istransient, since calcium returns to the cytoplasm through regulatedcalcium efflux channels, for instance a mitochondrial calcium antiportersuch as the mitochondrial calcium/sodium antiporter (MCA) also known asthe mitochondrial sodium/calcium exchanger (mNCE; see, e.g., Newgard1995; Magnus 1998; for a general review of mitochondrial membranetransporters, see, e.g., Zonatti et al., 1994 J. Bioenergetics Biomembr.26:543 and references cited therein). The use of MCA inhibitors has beencontemplated for their potential effects on cardiac function (e.g., Coxand Matlib, 1993 Trends Pharmacol. Sci. 14:408-413), but such use hasnot been suggested for certain other indications such as diabetes. Thus,for example, while elevated intramitochondrial calcium concentration hasbeen correlated with insulin secretion and oxidative ATP synthesis, asnoted above (e.g., Kennedy et al., 1996 J. Clin. Invest. 98:2524;Maechler et al., 1997 EMBO J. 16:3833; Cox and Matlib, 1993 TrendsPharmacol. Sci. 14:408), no inducer-effector relationship betweenoxidative ATP synthesis and insulin secretion has been universallyaccepted (see, e.g., Newgard, 1995 Ann. Rev. Biochem. 64:689). Moreover,currently available inhibitors of the MCA are regarded as either notspecific for the MCA, or useful only at extremely high concentrations,precluding their apparent suitability for pharmaceutical compositions(Cox and Matlib, 1993 Trends Pharmacol. Sci. 14:408-413).

Current pharmacological therapies for type 2 DM include injectedinsulin, and oral agents that are designed to lower blood glucoselevels. Currently available oral agents include: (i) the sulfonylureas,which act by enhancing the sensitivity of the pancreatic beta cell toglucose, thereby increasing insulin secretion in response to a givenglucose load; (ii) the biguanides, which improve glucose disposal ratesand inhibit hepatic glucose output; (iii) the thiazolidinediones, whichimprove peripheral insulin sensitivity through interaction with nuclearperoxisome proliferator-activated receptors (PPAR, see, e.g.,Spiegelman, 1998 Diabetes 47:507-514; Schoonjans et al., 1997 Curr.Opin. Lipidol. 8:159-166; Staels et al., 1997 Biochimie 79:95-99); (iv)repaglinide, which enhances insulin secretion through interaction withATP-dependent potassium channels; and (v) acarbose, which decreasesintestinal absorption of carbohydrates. Although currently availabledrugs for treating type 2 diabetes, such as the sulfonylureas, improveinsulin secretion, both basal and insulin stimulated insulin secretionare enhanced by such compounds. Consequently, undesirable chronichyperinsulinemia, hypoglycemia and/or excessive weight gain may resultfollowing treatment with such drugs (Cobb et al., 1998 Ann. Rep. Med.Chem. 33:213-222; Krentz et al., 1994 Drug Safety 11:223-241).

None of the current pharmacological therapies corrects the underlyingbiochemical defect in type 2 DM. Neither do any of these currentlyavailable treatments improve all of the physiological abnormalities intype 2 DM such as impaired insulin secretion, insulin resistance and/orexcessive hepatic glucose output. In addition, treatment failures arecommon with these agents, such that multi-drug therapy is frequentlynecessary.

Mitochondria are organelles that are the main energy source in cells ofhigher organisms. These organelles provide direct and indirectbiochemical regulation of a wide array of cellular respiratory,oxidative and metabolic processes, including metabolic energyproduction, aerobic respiration and intracellular calcium regulation.For example, mitochondria are the site of electron transport chain (ETC)activity, which drives oxidative phosphorylation to produce metabolicenergy in the form of adenosine triphosphate (ATP), and which alsounderlies a central mitochondrial role in intracellular calciumhomeostasis. These processes require the maintenance of a mitochondrialmembrane electrochemical potential, and defects in such membranepotential can result in a variety of disorders.

Mitochondria contain an outer mitochondrial membrane that serves as aninterface between the organelle and the cytosol, a highly folded innermitochondrial membrane that appears to form attachments to the outermembrane at multiple sites, and an intermembrane space between the twomitochondrial membranes. The subcompartment within the innermitochondrial membrane is commonly referred to as the mitochondrialmatrix (for review, see, e.g., Ernster et al., J. Cell Biol. 91:227s,1981). While the outer membrane is freely permeable to ionic andnon-ionic solutes having molecular weights less than about tenkilodaltons, the inner mitochondrial membrane exhibits selective andregulated permeability for many small molecules, including certaincations, and is impermeable to large (greater than about 10 kD)molecules.

Four of the five multisubunit protein complexes (Complexes I, III, IVand V) that mediate ETC activity are localized to the innermitochondrial membrane. The remaining ETC complex (Complex II) issituated in the matrix. In at least three distinct chemical reactionsknown to take place within the ETC, protons are moved from themitochondrial matrix, across the inner membrane, to the intermembranespace. This disequilibrium of charged species creates an electrochemicalmembrane potential of approximately 220 mV referred to as the“protonmotive force” (PMF). The PMF, which is often represented by thenotation Δp, corresponds to the sum of the electric potential (ΔΨm) andthe pH differential (ΔpH) across the inner membrane according to theequationΔp=Δm−ZΔpHwherein Z stands for −2.303 RT/F. The value of Z is −59 at 25° C. whenΔp and ΔΨm are expressed in mV and ΔpH is expressed in pH units (see,e.g., Ernster et al., J. Cell Biol. 91:227s, 1981, and references citedtherein).

ΔΨm provides the energy for phosphorylation of adenosine diphosphate(ADP) to yield ATP by ETC Complex V, a process that is coupledstoichiometrically with transport of a proton into the matrix. ΔΨm isalso the driving force for the influx of cytosolic Ca²⁺ into themitochondrion. Normal alterations of intramitochondrial Ca²⁺ areassociated with normal metabolic regulation (Dykens, 1998 inMitochondria & Free Radicals in Neurodegenerative Diseases, Beal, Howelland Bodis-Wollner, Eds., Wiley-Liss, New York, pp. 29-55; Radi et al.,1998 in Mitochondria & Free Radicals in Neurodegenerative Diseases,Beal, Howell and Bodis-Wollner, Eds., Wiley-Liss, New York, pp. 57-89;Gunter and Pfeiffer, 1991, Am. J. Physiol. 27: C755; Gunter et al., Am.J. Physiol. 267:313, 1994). For example, fluctuating levels ofmitochondrial free Ca²⁺ may be responsible for regulating oxidativemetabolism in response to increased ATP utilization, via allostericregulation of enzymes (reviewed by Crompton and Andreeva, Basic Res.Cardiol. 88:513-523, 1993), and the glycerophosphate shuttle (Gunter andGunter, J. Bioenerg. Biomembr. 26:471, 1994).

Normal mitochondrial function includes regulation of cytosolic freecalcium levels by sequestration of excess Ca²⁺ within the mitochondrialmatrix, including transiently elevated cytosolic free calcium thatresults from physiologic biological signal transduction. Depending oncell type, cytosolic Ca²⁺ concentration is typically 50-100 nM. Innormally functioning cells, when Ca²⁺ levels reach 200-300 nM,mitochondria begin to accumulate Ca²⁺ as a function of the equilibriumbetween influx via a Ca²⁺ uniporter in the inner mitochondrial membraneand Ca²⁺ efflux via both Na⁺ dependent and Na⁺ independent calciumcarriers, including notably the MCA. The low affinity of this rapiduniporter mechanism suggests that the primary uniporter function may beto lower cytosolic Ca²⁺ in response to elevation of cytosolic freecalcium levels, which may result from calcium influx across the plasmamembrane that occurs as part of a biological signal transductionmechanism (Gunter and Gunter, J. Bioenerg. Biomembr. 26:471, 1994;Gunter et al., Am. J. Physiol. 267:313, 1994). In certain instances, forexample in pancreatic beta cells, physiologic rises in cytoplasmiccalcium occur in response to glucose (or other secretagogues) and leadto calcium uptake by mitochondria, stimulating increased ATP synthesis.Similarly, the primary calcium antiporter (e.g., MCA) function may be tolower mitochondrial Ca²⁺ concentrations in response to mitochondrialCa²⁺ influxes, such as may result from glucose stimulation of aglucose-sensitive cell, and which produce transient increases inoxidative ATP synthesis. Thus, mitochondrially regulated calcium cyclingbetween, inter alia, cytosolic and mitochondrial compartments mayprovide an opportunity for manipulation of intracellular ATP levels(e.g., Cox and Matlib, 1993 Trends Pharmacol. Sci. 14:408-413; Matlib etal., 1983 Eur. J. Pharmacol. 89:327; Matlib 1985 J. Pharmacol. Exp.Therap. 233:376; Matlib et al. 1983 Life Sci. 32:2837).

In view of the significance of mitochondrial regulation of intracellularcalcium and the relationship of this mitochondrial activity to diabetes,which includes any of a wide range of disease states characterized byinappropriate and sustained hyperglycemia, there is clearly a need foragents to control mitochondrial calcium homeostasis. To provide improvedtherapies for diabetes, agents that alter mitochondrial calcium cyclingbetween intramitochondrial and extramitochondrial subcellularcompartments would be beneficial. Further, there is a need for improvedtherapeutics that are targeted to correct biochemical and/or metabolicdefects responsible for, or associated with, type 2 DM, regardless ofwhether such a defect underlying altered mitochondrial function may havemitochondrial or extramitochondrial origins.

Accordingly, there is a need in the art agents that modulatemitochondrial calcium/sodium antiporter function and are thus useful fortreating diabetes, type 2 DM, by enhancing insulin secretion. There isalso a need for pharmaceutical compositions containing such agents, aswell as for methods relating to use thereof. The present inventionfulfills these needs, and provides further related advantages.

SUMMARY OF THE INVENTION

In brief, this invention is generally directed to compounds that enhanceinsulin secretion, and thus are useful for the treatment of diabetesmellitus. Thus, in one embodiment, methods are disclosed for treatingdiabetes mellitus by administration a compound to a subject having orsuspected of being at risk for having diabetes mellitus, wherein thecompound has the following general structure (I):

including stereoisomers, prodrugs and pharmaceutically acceptable saltsthereof, wherein W₁, W₂, X, R₁, R₂, R₃, R₄, m and n are defined herein.

In one aspect of this embodiment, the diabetes mellitus is type 2diabetes mellitus or maturity onset diabetes of the young. In anotheraspect, the compounds enhance insulin secretion, such as insulinsecretion that is stimulated by glucose. In other aspects, the compoundsenhances insulin secretion that is stimulated by a supraphysiologicalglucose concentration, and does not enhance insulin secretion in thepresence of a physiological glucose concentration. In further aspects,the methods may further comprise administering to the subject one ormore agents that lowers circulating glucose concentration in thesubject, such as insulin, an insulin secretagogue, an insulinsensitizer, an inhibitor of hepatic glucose output and/or an agent thatimpairs glucose absorption.

In other embodiments, pharmaceutical compositions are disclosed thatcontain one or more compounds having structure (I) in combination withone or more pharmaceutically acceptable carriers, as well as novelcompounds within structure (1).

These and other aspects of the present invention will become apparentupon reference to the following detailed description and attacheddrawings. All references disclosed herein are hereby incorporated byreference in their entireties as if each was incorporated individually.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1 and 2 shows enhanced glucose stimulated insulin secretion byINS-1 cells (FIG. 1) and on rat pancreatic islet cells (FIG. 2) whenexposed to CPG37157, a known potent inhibitor of MCA, in the presence ofbasal or supraphysiological glucose.

FIG. 3 shows blood glucose concentrations following a glucose load attime 0, in db/db mutant mice treated one hour prior to time 0 withcompound no. 16o (▴) or with vehicle only (▪); (error bars show SEM,*p<0.05).

DETAILED DESCRIPTION OF THE INVENTION

This invention is generally directed to compounds that enhance insulinsecretion, and thus are useful for the treatment of diabetes mellitus.Thus, in one embodiment, methods are disclosed for treating diabetesmellitus by administration a compound to a subject having or suspectedof being at risk for having diabetes mellitus, wherein the compounds hasthe following general structure (I):

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof,

-   -   wherein        -   X is —S(O)_(q)—, —O—, —N(R)— or —C(R)(R′)—;        -   m is 0 or 1;        -   n is 0, 1 or 2;        -   q is 0, 1 or 2;        -   W₁ and W₂ each represent an optional substituent, wherein W₁            and W₂ are the same or different and independently halogen,            nitro, or lower alkyl;        -   R and R′ are the same or different and independently alkyl,            substituted alkyl, aryl, substituted aryl, arylalkyl or            substituted arylalkyl,        -   or R and R′ taken together with the carbon atom to which            they are bonded form a carbocycle, substituted carbocycle,            heterocycle or substituted heterocycle;        -   R₁ is alkyl, substituted alkyl, aryl, substituted aryl,            arylalkyl, substituted arylalkyl, heteroaryl or substituted            heteroaryl;        -   R₂ is hydrogen, nitro, —OR_(2a), —C(═O)NR_(2b)R_(2c),            —CH₂NR_(2b)R_(2c), —CH₂OR_(2a), —NR_(2b)R_(2c),            —NHC(═O)R_(2a), —NHC(═O)NR_(2b)R_(2c) or            —NHC(═NH)NR_(2b)R_(2c);        -   R_(2a) is hydrogen, alkyl, substituted alkyl, arylalkyl, or            substituted arylalkyl;        -   R_(2b) and R_(2c) are the same or different and            independently hydrogen, alkyl, substituted alkyl, —SO₂R₄,            —C(═NH)NH₂ or —C(═O)R_(2d) where R_(2d) is amino, alkyl,            substituted alkyl, aryl or substituted aryl;        -   R₃ is hydroxy, alkyl, substituted alkyl, aryl, substituted            aryl, arylalkyl, substituted arylalkyl, heterocycle,            substituted heterocycle, heterocyclealkyl, substituted            heterocyclealkyl, —C(═O)N(R_(3a))(R_(3b)),            —NHC(═O)N(R_(3a))(R_(3b)), —NHC(═S)N(R_(3a))(R_(3b)),            —C(═O)OR_(3c), —C(═O)R_(3c), —NHC(═O)R_(3d) or —NHSO₂R_(3d);        -   R_(3a) and R_(3b) are the same or different and            independently hydrogen, alkyl, substituted alkyl, aryl,            substituted aryl, arylalkyl or substituted arylalkyl,        -   or R_(3a) and R_(3b) taken together with the nitrogen atom            to which they are attached form a heterocycle or substituted            heterocycle;        -   R_(3c) is hydrogen, alkyl, substituted alkyl, aryl,            substituted aryl, arylalkyl, substituted arylalkyl,            heteroaryl, substituted heteroaryl, heteroarylalkyl or            substituted heteroarylalkyl;        -   R_(3d) is alkyl, substituted alkyl, aryl, substituted aryl,            arylalkyl, substituted arylalkyl, heteroaryl, substituted            heteroaryl, heteroarylalkyl or substituted heteroarylalkyl;            and        -   R₄ is, at each occurrence, the same or different and            independently hydrogen, alkyl, substituted alkyl, aryl,            substituted aryl, arylalkyl or substituted arylalkyl.

As used herein, the terms used above have the following meaning:

“Alkyl” means a straight chain or branched, saturated or unsaturated,cyclic or non-cyclic hydrocarbon having from 1 to 10 carbon atoms, while“lower alkyl” has the same meaning but only has from 1 to 6 carbonatoms. Representative saturated straight chain alkyls include methyl,ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, and the like; whilesaturated branched alkyls include isopropyl, sec-butyl, isobutyl,tert-butyl, isopentyl, and the like. Unsaturated alkyls contain at leastone double or triple bond between adjacent carbon atoms (also referredto as an “alkenyl” or “alkynyl”, respectively). Representative straightchain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl,2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; whilerepresentative straight chain and branched alkynyls include acetylenyl,propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1butynyl, and the like. Representative saturated cyclic alkyls includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂cyclohexyl and thelike; while unsaturated cyclic alkyls include cyclopentenyl,cyclohexenyl, —CH₂cyclohexenyl and the like. Cycloalkyls are alsoreferred to herein as “carbocyclic” rings systems, and include bi- andtri-cyclic ring systems having from 8 to 14 carbon atoms such as acycloalkyl (such as cyclopentane or cyclohexane) fused to one or morearomatic (such as phenyl) or non-aromatic (such as cyclohexane)carbocyclic rings.

“Halogen” means fluorine, chlorine, bromine or iodine.

“Oxo” means a carbonyl group (i.e., ═O).

“Mono- or di-alkylamino represents an amino substituted with one alkylor with two alkyls, respectively.

“Alkanediyl” means a divalent alkyl from which two hydrogen atoms aretaken from the same carbon atom or from different carbon atoms, such as—CH₂— —CH₂CH₂—, —CH₂CH₂CH₂—, —CH(CH₃)CH₂—, and the like.

“Aryl” means an aromatic carbocyclic moiety such as phenyl or naphthyl.

“Arylalkyl” means an alkyl having at least one alkyl hydrogen atomreplaced with an aryl moiety, such as benzyl, —(CH₂)₂phenyl,—(CH₂)₃phenyl, —CH(phenyl)₂, and the like.

“Heteroaryl” means an aromatic heterocycle ring of 5 to 10 members andhaving at least one heteroatom selected from nitrogen, oxygen andsulfur, and containing at least 1 carbon atom, including both mono- andbicyclic ring systems. Representative heteroaryls are pyridyl, furyl,benzofuranyl, thiophenyl, benzothiophenyl, quinolinyl, pyrrolyl,indolyl, oxazolyl, benzoxazolyl, imidazolyl, benzimidazolyl, thiazolyl,benzothiazolyl, isoxazolyl, pyrazolyl, isothiazolyl, pyridazinyl,pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, andquinazolinyl.

“Heteroarylalkyl” means an alkyl having at least one alkyl hydrogen atomreplaced with a heteroaryl moiety, such as —CH₂pyridinyl,—CH₂pyrimidinyl, and the like.

“Heterocycle” means a 5- to 7-membered monocyclic, or 7- to 10-memberedbicyclic, heterocyclic ring which is either saturated, unsaturated, oraromatic, and which contains from 1 to 4 heteroatoms independentlyselected from nitrogen, oxygen and sulfur, and wherein the nitrogen andsulfur heteroatoms may be optionally oxidized, and the nitrogenheteroatom may be optionally quaternized, including bicyclic rings inwhich any of the above heterocycles are fused to a benzene ring. Theheterocycle may be attached via any heteroatom or carbon atom.Heterocycles include heteroaryls as defined above. Thus, in addition tothe heteroaryls listed above, heterocycles also include morpholinyl,pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl,valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl,tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

“Heterocyclealkyl” means an alkyl having at least one alkyl hydrogenatom replaced with a heterocycle, such as —CH₂morpholinyl, and the like.

The term “substituted” as used herein means any of the above groups(i.e., alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycleand heterocyclealkyl) wherein at least one hydrogen atom is replacedwith a substituent (also referenced herein as “Q”). In the case of anoxo substituent (“═O”) two hydrogen atoms are replaced. Substituentsinclude halogen, hydroxy, oxo, alkyl, substituted alkyl (such ashaloalkyl, mono- or di-substituted aminoalkyl, alkyloxyalkyl, and thelike), aryl, substituted aryl, arylalkyl, substituted arylalkyl,heteroaryl, substituted heteroaryl, heteroarylalkyl, substitutedheteroarylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl,substituted heterocyclealkyl, —NR_(a)R_(b), —NR_(a)C(═O)R_(b),—NR_(c)C(═O)NR_(a)R_(b), —NR_(a)C(═O)OR_(b), —NR_(a)SO₂R_(b), —OR_(a),—C(═O)R_(a), —C(═O)OR_(a), —C(═O)NR_(a)R_(b), —OC(═O)R_(a),—OC(═O)OR_(a), —OC(═O)NR_(a)R_(b), —NR_(a)SO₂R_(b),—CONR_(a)(alkanediyl)OR_(b),—CONR_(c)(alkanediyl-O)₁₋₆(alkanediyl)NR_(a)R_(b), or a radical of theformula —Y—Z—R_(a) where Y is alkanediyl, substituted alkanediyl or adirect bond, Z is —O—, —S—, —S(═O)—, —S(═O)₂—, —N(R_(b))—, —C(═O)—,—C(═O)O—, —OC(═O)—, —N(R_(b))C(═O)—, —C(═O)N(R_(b))— or a direct bond,wherein R_(a), R_(b) and R_(c) are the same or different andindependently hydrogen, amino, alkyl, substituted alkyl (includinghalogenated alkyl), aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl,substituted heteroarylalkyl, heterocycle, substituted heterocycle,heterocyclealkyl or substituted heterocyclealkyl, or wherein R_(a) andR_(b) taken together with the nitrogen atom to which they are attachedform a heterocycle or substituted heterocycle.

“Pharmaceutically acceptable salt” refers to salts of the compounds ofthe present invention derived from the combination of such compounds andan organic or inorganic acid (acid addition salts) or an organic orinorganic base (base addition salts). The compounds of the presentinvention may be used in either the free base or salt forms, with bothforms being considered as being within the scope of the presentinvention.

The compounds of the present invention may generally be utilized as thefree acid or base. Alternatively, the compounds of this invention may beused in the form of acid or based addition salts. Acid addition salts ofthe free base amino compounds of the present invention may be preparedby methods well known in the art, and may be formed from organic andinorganic acids. Suitable organic acids include maleic, fumaric,benzoic, ascorbic, succinic, methanesulfonic, acetic, oxalic, propionic,tartaric, salicylic, citric, gluconic, lactic, mandelic, cinnamic,aspartic, stearic, palmitic, glycolic, glutamic, and benzenesulfonicacids. Suitable inorganic acids include hydrochloric, hydrobromic,sulfuric, phosphoric, and nitric acids. Based addition salts include theammonium ion, as well as other suitable cations. Thus, the term“pharmaceutically acceptable salt” of structure (I) is intended toencompass any and all acceptable salt forms.

In addition, prodrugs are also included within the context of thisinvention. Prodrugs are any covalently bonded carriers that release acompound of structure (I) in vivo when such prodrug is administered to apatient. Prodrugs are generally prepared by modifying functional groupsin a way such that the modification is cleaved, either by routinemanipulation or in vivo, yielding the parent compound.

With regard to stereoisomers, the compounds of structure (I) may havechiral centers and may occur as racemates, racemic mixtures and asindividual enantiomers or diastereomers. All such isomeric forms areincluded within the present invention, including mixtures thereof.Furthermore, some of the crystalline forms of the compounds of structure(1) may exist as polymorphs, which are included in the presentinvention. In addition, some of the compounds of structure (I) may alsoform solvates with water or other organic solvents. Such solvates aresimilarly included within the scope of this invention.

The compounds of structure (I), as well as the more specific embodimentsdiscussed below, may be made by techniques knows to those skilled in thefield of organic chemistry, and as more specifically exemplified in theExamples.

In one embodiment, X is —S(O)_(q), and the compounds have the followingstructure (II):

In one aspect of this embodiment, q is 2, 1 or 0 and the compounds havethe following structure (II-1), (II-2) or (II-3), respectively:

In another embodiment, X is —O— and the compounds have the followingstructure (III):

In a further embodiment, X is —N(R)— and the compounds have thefollowing structure (IV):

In yet another embodiment, X is C(R)(R′)— and the compounds have thefollowing structure (V):

Depending upon the choice of m in structure (I), the compound have thefollowing structure (I-1) when m is 0 and structure (I-2) when m is 1:

Depending upon the choice of n, compounds of this invention have thefollowing structure (I-3) when n is 0, structure (14) when n is 1, andstructure (I-5) when n is 2:

Depending upon the choice of the R₂ group, compounds have the followingstructure (I-6) when R₂ is hydrogen, structure (I-7) when R₂ is —OR_(e),structure (I-8) when R₂ is —C(═O)NR_(2b)R_(2c), structure (I-9) when R₂is —CH₂NR_(2b)R_(2c), structure (I-10) when R₂ is —CH₂OR_(2a), structure(I-11) when R₂ is —NR_(2b)R_(2c), structure (I-12) when R₂ is—NHC(═O)R_(2a), structure (I-13) when R₂ is —NHC(═O)NR_(2b)R_(2c), andstructure (I-14) when R₂ is —NHC(═NH)NR_(2b)R_(2c):

In one embodiment, R₁ is aryl or substituted aryl and the compounds havethe following structure (VI):

wherein Ar represents aryl or substituted aryl as defined herein.

In a more specific aspect of structure (VI), W₁ is present at the5-position, W₂ is not present, and the compounds have the followingstructure (VII):

In a particular embodiment of structure (VII), W₁ is halogen, such aschloro, bromo or fluoro, and more particularly chloro.

In more specific embodiments of structure (VII), X is S(O)_(q) where qis 0, R₂ is —NR_(2a)R_(2c), n is 1 and m is 0 or 1, and the compoundshave the following structure (VII-1) or (VII-2), respectively:

In other more specific embodiments of structure (VII), X is S(O)_(q)where q is 0, R₂ is —NR_(2a)R_(2c), n is 2 and m is 0 or 1, and thecompounds have the following structure (VII-3) or (VI-4), respectively:

In another embodiment, R₁ is alkyl or substituted alkyl and thecompounds have the following structure (VI):

wherein Alk represents alkyl or substituted alkyl as defined herein.

In a more specific aspect of structure (VIII), W₁ is present at the5-position, W₂ is not present, and the compounds have the followingstructure (IX):

In a particular embodiment of structure (IX), W₁ is halogen, such aschloro, bromo or fluoro, and more particularly chloro.

In more specific embodiments of structure (IX), X is S(O)_(q) where q is0, R₂ is —NR_(2a)R_(2c), n is I and m is 0 or 1, and the compounds havethe following structure (IX-1) or (IX-2), respectively:

In more specific embodiments of structure (IX), X is S(O)_(q) where q is0, R₂ is —NR_(2a)R_(2c), n is 2 and m is 0 or 1, and the compounds havethe following structure (IX-3) or (IX-4), respectively:

In other specific embodiments, X is S(O)_(q), R₃ is—C(═O)N(R_(3a))(R_(3b)), —C(═O)OR_(3c), —C(═O)R_(3c), heterocycle orsubstituted heterocycle, —NHC(═O)N(R_(3a))(R_(3b)),—NHC(═S)N(R_(3a))(R_(3b)), —NHC(═O)R_(3d) or —NHSO₂R_(3d), R₄ ishydrogen, and the compounds have the following structures (X) through(XVII), respectively:

Although not intending to be limited by the following theory, it isbelieved that compounds of structure (I) are selective inhibitors of themitochondrial calcium/sodium antiporter (MCA). As described in greaterdetail herein, such compounds substantially enhance insulin secretion.In one aspect, the compounds enhance insulin secretion that isstimulated by supraphysiological glucose concentrations (e.g., glucosestimulated insulin secretion), but does not substantially enhanceinsulin secretion under conditions where normal physiological glucoseconcentrations are present (e.g., basal insulin secretion). In thisaspect of the invention, the compounds selectively interfere with MCAand/or other mitochondrial calcium efflux mechanisms in a manner thatpreferentially enhances glucose stimulated insulin secretion relative tobasal insulin secretion, and thus are particularly useful for treatmentof diabetes.

More specifically, it is believed that the compounds of structure (1)maintain increased and sustained intramitochondrial calciumconcentrations, thereby driving oxidative phosphorylation and theconsequent elevation of intracellular ATP concentration. Such elevatedATP concentrations promote enhanced insulin secretion and effect thedesirable result of providing sufficient insulin to lowersupraphysiological circulating glucose concentrations and preferablyreturn them to concentrations at or near normal levels.

In certain aspects, there is provided a method for treating diabetescomprising administering to a subject a therapeutically effective amountof a compound of structure (I), and further comprising administering anagent that lowers circulating glucose concentrations. While currentagents for treating type 2 DM may lower blood glucose levels withoutcorrecting underlying biochemical defects in this disease, it isdesirable in certain instances to combine a compound of structure (I)with an existing hypoglycemic agent. For example, an agent of thesulfonylurea class or of the more recently developed non-sulfonylureaclass of agents that close the potassium/ATP channel may be combinedwith a compound of stucture (I). As other non-limiting examples, agentsthat supply substrates for mitochondrial metabolism (e.g., KCl,α-ketoisocaproic acid or leucine), insulin sensitizers (e.g.,thiazolidinediones), inhibitors of hepatic glucose output (e.g.,metformin) or glucose uptake blockers (e.g., acarbose) may also beemployed.

An “agent that lowers circulating glucose concentrations” includes anyhypoglycemic agent as known in the art and provided herein, includinganti-diabetic agents such as sulfonylurea compounds and non-sulfonylureacompounds, and may further include a biguanide, a thiazolidinedione,repaglinide, acarbose, metformin or other hypoglycemic compositions(e.g., 6LP-1 and its analogs, DPP-IV inhibitors, α-ketoisocaproic acid,leucine or analogs of other amino acids).

A “biological sample” may comprise any tissue or cell preparation asdescribed herein and a “biological sample containing a mitochondrialcalcium/sodium antiporter polypeptide” comprises any tissue or cellpreparation in which an expressed MCA polypeptide or other mitochondrialmolecular component as provided herein that mediates Ca²⁺ efflux from amitochondrion is thought to be present. Biological samples (includingthose containing a MCA polypeptide) may be provided by obtaining a bloodsample, biopsy specimen, tissue explant, organ culture or any othertissue or cell preparation from a subject or a biological source. Thesubject or biological source may be a human or non-human animal, aprimary cell culture or culture adapted cell line including but notlimited to genetically engineered cell lines that may containchromosomally integrated or episomal recombinant nucleic acid sequences,immortalized or immortalizable cell lines, somatic cell hybrid orcytoplasmic hybrid “cybrid” cell lines, differentiated ordifferentiatable cell lines, transformed cell lines and the like. Abiological sample may, for example, be derived from a recombinant cellline or from a transgenic animal.

In certain preferred embodiments the subject or biological source is ahuman known to have, or suspected of being at risk for having, diabetesmellitus. In certain further preferred embodiments the diabetes mellitusis type 2 diabetes mellitus, and in certain other further preferredembodiments the diabetes mellitus is maturity onset diabetes of theyoung (MODY). Well known criteria have been established for determininga presence of, or risk for having diabetes mellitus (e.g., type 2diabetes mellitus, MODY) as described herein and as known in the art,and these may be found, for example, in Clinical PracticeRecommendations 2000 (2000 Diabetes Care 23: supplement 1) or elsewhere(see, e.g., www.diabetes.org/, the website of the American DiabetesAssociation). Among these recognized physiological parameters thatrelate to diabetes, those familiar with the art will appreciate that avariety of methodologies have been established for the determination ofglucose and insulin concentrations in the circulation. For example,methods for quantifying insulin in a biological sample as providedherein (e.g., a blood, serum or plasma sample) may include aradioimmunoassay (RIA) using an antibody that specifically binds toinsulin. Variations on RIA such as enzyme linked immunosorbent assaysand immunoprecipitation analysis, and other assays for the presence ofinsulin or proinsulin in a biological sample are readily apparent tothose familiar with the art, and may further include assays that measureinsulin secretion by cells in the presence or absence of secretagoguessuch as glucose, KCl, amino acids, sulfonylureas, forskolin,glyceraldehyde, succinate or other agents that may increase or decreaseinsulin or proinsulin in a cell conditioned medium. Such methods mayalso be used to quantify the amount of insulin produced by or releasedfrom an insulin-secreting cell.

Because it is well recognized by those familiar with the art that theremay be large quantitative variations in circulating glucose and insulinlevels among individual subjects (see, e.g., Clinical PracticeRecommendations 2000, 2000 Diabetes Care 23 (suppl. 1), and referencescited therein), the present invention contemplates in preferredembodiments a method for treating diabetes with a pharmaceuticalcomposition comprising a compound of structure (I) that selectivelyimpairs MCA activity as provided herein, wherein the compound does notsubstantially enhance insulin secretion at physiological glucoseconcentration (i.e., under fasting or basal metabolic conditions) andwherein the compound substantially enhances insulin secretion atsupraphysiological glucose concentration (i.e., under non-fastingconditions or conditions of glucose stimulation). Although certainpreferred embodiments of the present invention relate to compositionsand methods for treating diabetes in humans, the invention need not beso limited. In particular, those having ordinary skill in the art willreadily appreciate that diabetes, including any disease statecharacterized by inappropriate and/or sustained periods of hyperglycemiasuch as type 2 DM or other diabetes mellitus, may be a condition that ispresent in a number of non-human animals (e.g., Ford, 1995 Veterin.Clinics of N. Amer.: Small Animal Practice 25(3):599-615). Accordingly,compositions and methods provided herein as may be useful for thetreatment of these and other manifestations of diabetes in non-humananimals are within the scope and spirit of the present invention.

Normal or fasting physiological glucose concentration thus refers to theconcentration of glucose in the circulation of a subject under normalconditions (e.g., fasting basal conditions), which are distinct fromtransient supraphysiological, non-fasting or otherwise temporarilyelevated glucose concentrations that are achieved under non-normalconditions such as after feeding or other conditions of glucosestimulation. For example by way of illustration and not limitation,depending on a variety of factors such as the physiological status,diet, activity level, health and/or genetic constitution of a subject,or the like, metabolic homeostatic mechanisms (including insulinsecretion) typically operate to maintain a relatively narrow range ofcirculating glucose concentrations under fasting conditions that aresignificantly lower than circulating glucose concentrations that arereached following feeding or other glucose stimulation. Such elevatedglucose concentrations, which typically are not sustained over time,reflect a departure from the normal or fasting state sought to bemaintained by the homeostatic mechanisms, and are referred to herein assupraphysiological glucose concentrations. Accordingly, and as a furthernon-limiting example, many normal individuals may maintain a fasting orphysiological circulating glucose concentration at or aroundapproximately 40-80 mg/dl and generally less than about 10 mg/dl, whichmay be generally less than 126 mg/dl in an individual characterized ashaving “impaired fasting glucose”, and which may be generally greaterthan 126 mg/dl in an individual characterized as diabetic (see, e.g.,Gavin et al., 2000 Diabetes Care 23 (suppl. 1):S4-S 19 and referencescited therein) such that a glucose concentration induced by feeding orother type of glucose stimulation that is greater than such a fasting orphysiological glucose concentration in a statistically significantmanner may be regarded as a supraphysiological glucose concentration.Similarly, there may be large variations among individuals with regardto circulating insulin concentrations and the degree to which an agentthat impairs MCA activity according to the invention effects elevatedinsulin concentrations. Therefore, the present invention contemplates“enhanced” insulin secretion to refer to an insulin concentration thatis, in a statistically significant manner, detectably increased by anMCA activity-impairing agent to a greater degree followingsupraphysiological glucose stimulation than is the degree (if any) towhich the MCA activity-impairing agent increases the detectable insulinconcentration under fasting or physiological conditions. Accordingly, inpreferred embodiments, the compound that selectively impairs an MCAactivity enhances insulin secretion that is stimulated by asupraphysiological glucose concentration and does not enhance insulinsecretion in the presence of a fasting glucose concentration.

It is important to an understanding of the present invention to notethat all technical and scientific terms used herein, unless otherwisedefined, are intended to have the same meaning as commonly understood byone of ordinary skill in the art. The techniques employed herein arealso those that are known to one of ordinary skill in the art, unlessstated otherwise. Throughout this application various publications arereferenced within parentheses. The disclosures of these publications intheir entireties are hereby incorporated by reference in thisapplication.

Reference to particular buffers, media, reagents, cells, cultureconditions and the like, or to some subclass of the same, is notintended to be limiting, but should be read to include all such relatedmaterials that one of ordinary skill in the art would recognize as beingof interest or value in the particular context in which that discussionis presented. For example, it is often possible to substitute one buffersystem or culture medium for another, such that a different but knownway is used to achieve the same goals as those to which the use of asuggested method, material or composition is directed.

According to certain embodiments of the present invention a“therapeutically effective amount” of a compound of structure (I) thatimpairs a MCA activity and/or a compound that lowers circulating glucoseconcentration may be administered. The person having ordinary skill inthe art can readily and without undue experimentation determine what isa therapeutically effective amount as provided herein. Thus, for exampleand as described elsewhere herein, in the context of diabetes, and morespecifically in the context of monitoring efficacy of diabetes therapy,periodic determination of circulating blood glucose concentrations maybe routinely performed in order to determine whether a subject's bloodglucose has attained a normal, physiological level. (see, e.g., Gavin etal., 2000 Diabetes Care 23 (suppl. 1):S4-S 19 and references citedtherein) Optionally or additionally, according to certain contemplatedembodiments it may be desirable to monitor blood insulin and/or glycatedhemoglobin levels, which as described herein may be performed accordingto any of a number of routine and well established methodologies.

Those having ordinary skill in the art are readily able to compare ATPproduction by an ATP biosynthetic pathway in the presence and absence ofa candidate ATP biosynthesis factor. Routine determination of ATPproduction may be accomplished using any known method for quantitativeATP detection, for example by way of illustration and not limitation, bydifferential extraction from a sample optionally includingchromatographic isolation; by spectrophotometry; by quantification oflabeled ATP recovered from a sample contacted with a suitable form of adetectably labeled ATP precursor molecule such as, for example, ³²P; byquantification of an enzyme activity associated with ATP synthesis ordegradation; or by other techniques that are known in the art.Accordingly, in certain embodiments of the present invention, the amountof ATP in a biological sample or the production of ATP (including therate of ATP production) in a biological sample may be an indicator ofaltered mitochondrial function. In one embodiment, for instance, ATP maybe quantified by measuring luminescence of luciferase catalyzedoxidation of D-luciferin, an ATP dependent process.

As described herein, a compound that selectively impairs MCA activitymay in certain preferred embodiments interfere with transmembranetransport of calcium cations, whereby such activity may be determined bydetecting calcium. A variety of calcium indicators are known in the artand are suitable for generating a detectable signal in solution or as anintracellular signal, for example, a signal that is proportional to thelevel of calcium in the cytosol, including but not limited tofluorescent indicators such as fura-2 (McCormack et al., 1989 Biochim.Biophys. Acta 973:420); mag-fura-2; BTC (U.S. Pat. No. 5,501,980);fluo-3, fluo-4, fluo-5F and fluo-5N (U.S. Pat. No. 5,049,673); fura-4F,fura-5F, fura-6F, and fura-FF; rhod-2 and rhod-5F; Calcium Green 5N™;benzothiaza-1 and benzothiaza-2; and others, which are available fromMolecular Probes, Inc., Eugene, Oreg. (see also, e.g., Calcium SignalingProtocols—Meths. In Mol. Biol.—Vol. 114, Lambert, D. (ed.), HumanaPress, 1999).

Calcium Green 5Nm is a particularly preferred calcium indicator moleculefor use according to the present invention. Depending, however, on theparticular assay conditions to be used, a person having ordinary skillin the art can select a suitable calcium indicator from those describedabove or from other calcium indicators, according to the teachingsherein and based on known properties (e.g., solubility, stability, etc.)of such indicators. For example by way of illustration and notlimitation, whether a cell permeant or cell impermeant indicator isneeded (e.g., whether a sample comprises a permeabilized cell), affinityof the indicator for calcium (e.g., dynamic working range of calciumconcentrations within a sample as provided herein) and/or fluorescencespectral properties such as a calcium-dependent fluorescence excitationshift, may all be factors in the selection of a suitable calciumindicator. Calcium-Green-5N™ (potassium salt) is commercially available(Molecular Probes, Eugene, Oreg.; C-3737). Calcium-Green-5N™ is a lowaffinity Ca²⁺ indicator (as is, for example, Oregon Green 488 BAPTA-5N).Low affinity indicators are preferred because of the Ca²⁺ concentrationsused in the assays. High affinity dyes require a lower Ca²⁺concentration and therefore a lower number of cells, and thus a lowernumber of mitochondria, would be required than the number used in theassays.

Other calcium-sensitive detectable reagents that can be used in theassay of the invention include Calcein, Calcein Blue, Calcium-Green-1,Calcium-Green-2, Calcium-Green-C₁₈, Calcium Orange, Calcium-Orange-5N,Calcium Crimson, Fluo-3, Fluo-3 AM ester, Fluo-4, Fura-2, Fura-2FF, FuraRed, Pura-Cl₈, Indo-1, Bis-Fura-2, Mag-Fura-2, Mag-Fura-5, Mag-Indo-1,Magnesium Green, Quin-2, Quin-2 AM (acetoxymethyl) ester, MethoxyquinMF, Methoxyquin MF AM ester, Rhod-2, Rhod-2 AM ester, Texas Red-CalciumGreen, Oregon Green 488 BAPTA-1, Oregon Green 488 BAPTA-2, BTC, BTC AMester, (all from Molecular probes, OR), and aequorin. As noted above, incertain preferred embodiments intramitochondrial calcium concentrationsare directly determined using mitochondrially targeted aequorin.

In the practice of the methods of this invention, compounds of structure(I) are typically administered to a patient in the form of apharmaceutically acceptable composition, which comprises one or morecompounds of structure (I) in combination with one or morepharmaceutically acceptable carrier(s). A “pharmaceutically acceptablecarrier” for therapeutic use are well known in the pharmaceutical art,and are described, for example, in Remingtons Pharmaceutical Sciences,Mack Publishing Co. (A. R. Gennaro edit. 1985). For example, sterilesaline and phosphate-buffered saline at physiological pH may be used.Preservatives, stabilizers, dyes and even flavoring agents may beprovided in the pharmaceutical composition. For example, sodiumbenzoate, sorbic acid and esters of p-hydroxybenzoic acid may be addedas preservatives. Id. at 1449. In addition, antioxidants and suspendingagents may be used. Id.

The pharmaceutical compositions that contain one or more compounds asprovided herein may be in any form which allows for the composition tobe administered to a patient. For example, the composition may be in theform of a solid, liquid or gas (aerosol). Typical routes ofadministration include, without limitation, oral, topical, parenteral(e.g., sublingually or buccally), sublingual, rectal, vaginal, andintranasal. The term parenteral as used herein includes subcutaneousinjections, intravenous, intramuscular, intrasternal, intracavernous,intrameatal, intraurethral injection or infusion techniques. Thepharmaceutical composition is formulated so as to allow the activeingredients contained therein to be bioavailable upon administration ofthe composition to a patient. Compositions that will be administered toa patient take the form of one or more dosage units, where for example,a tablet may be a single dosage unit, and a container of one or morecompounds of the invention in aerosol form may hold a plurality ofdosage units.

For oral administration, which is the route of administration inpreferred embodiments, an excipient and/or binder may be present.Examples are sucrose, kaolin, glycerin, starch dextrins, sodiumalginate, carboxymethylcellulose and ethyl cellulose. Coloring and/orflavoring agents may be present. A coating shell may be employed.

The composition may be in the form of a liquid, e.g., an elixir, syrup,solution, emulsion or suspension. The liquid may be for oraladministration or for delivery by injection, as two examples. Whenintended for oral administration, preferred compositions contain, inaddition to one or more compounds of structure (I), one or more of asweetening agent, preservatives, dye/colorant and flavor enhancer. In acomposition intended to be administered by injection, one or more of asurfactant, preservative, wetting agent, dispersing agent, suspendingagent, buffer, stabilizer and isotonic agent may be included.

A liquid pharmaceutical composition as used herein, whether in the formof a solution, suspension or other like form, may include one or more ofthe following adjuvants: sterile diluents such as water for injection,saline solution, preferably physiological saline, Ringer's solution,isotonic sodium chloride, fixed oils such as synthetic mono ordiglycerides which may serve as the solvent or suspending medium,polyethylene glycols, glycerin, propylene glycol or other solvents;antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parenteral preparation can be enclosedin ampoules, disposable syringes or multiple dose vials made of glass orplastic. Physiological saline is a preferred adjuvant. An injectablepharmaceutical composition is preferably sterile.

A liquid composition intended for either parenteral or oraladministration should contain an amount of a compound as provided hereinsuch that a suitable dosage will be obtained. Typically, this amount isat least 0.01 wt % of the compound in the composition. When intended fororal administration, this amount may be varied to be between 0.1 andabout 70% of the weight of the composition. Preferred oral compositionscontain between about 4% and about 50% of the compound(s). Preferredcompositions and preparations are prepared so that a parenteral dosageunit contains between 0.01 to 1% by weight of the compound.

The pharmaceutical composition may be intended for topicaladministration, in which case the carrier may suitably comprise asolution, emulsion, ointment or gel base. The base, for example, maycomprise one or more of the following: petrolatum, lanolin, polyethyleneglycols, beeswax, mineral oil, diluents such as water and alcohol, andemulsifiers and stabilizers. Thickening agents may be present in apharmaceutical composition for topical administration. If intended fortransdermal administration, the composition may include a transdermalpatch or iontophoresis device. Topical formulations may contain aconcentration of the compound of from about 0.1 to about 10% w/v (weightper unit volume).

The composition may be intended for rectal administration, in the form,e.g., of a suppository that will melt in the rectum and release thedrug. The composition for rectal administration may contain anoleaginous base as a suitable nonirritating excipient. Such basesinclude, without limitation, lanolin, cocoa butter and polyethyleneglycol. In the methods of the invention, the agent(s) that altermitochondrial function identified as described herein may beadministered through use of insert(s), bead(s), timed-releaseformulation(s), patch(es) or fast-release formulation(s).

It will be evident to those of ordinary skill in the art that theoptimal dosage of the compound(s) may depend on the weight and physicalcondition of the patient; on the severity and longevity of the physicalcondition being treated; on the particular form of the activeingredient, the manner of administration and the composition employed.The use of the minimum dosage that is sufficient to provide effectivetherapy is usually preferred. Patients may generally be monitored fortherapeutic or prophylactic effectiveness using assays suitable for thecondition being treated or prevented, which will be familiar to thosehaving ordinary skill in the art and which, as noted above, willtypically involve determination of whether circulating insulin and/orglucose concentrations fall within acceptable parameters according towell-known techniques. Suitable dose sizes will vary with the size,condition and metabolism of the patient, but will typically range fromabout 10 mL to about 500 mL for 10-60 kg individual. It is to beunderstood that according to certain embodiments the compound may bemembrane permeable, preferably permeable through the plasma membraneand/or through mitochondrial outer and/or inner membranes. According tocertain other embodiments, the use of the compound as disclosed hereinin a chemotherapeutic composition can involve such an agent being boundto another compound, for example, a monoclonal or polyclonal antibody, aprotein or a liposome, which assist the delivery of said compound.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Although the foregoing invention has beendescribed in some detail by way of illustration and example for purposesof clarity of understanding, it will be readily apparent to those ofordinary skill in the art in light of the teachings of this inventionthat certain changes and modifications may be made thereto withoutdeparting from the spirit or scope of the appended claims.

The following Examples illustrate the invention and are not intended tolimit the same. Those skilled in the art will recognize, or be able toascertain through routine experimentation, numerous equivalents to thespecific substances and procedures described herein. Such equivalentsare considered to be within the scope of the present invention.

EXAMPLE 1

Synthesis of 4-chloro-1′,1′,1′-trimethylacetanilide (2a)

To a solution of 4-chloroaniline (1a) (20 g, 0.157 mole) in DCM (400 ml)at 0° C. was added DIEA (54.7 ml, 0.314 mol) and pivaloyl chloride (23.2ml, 0.188 mol). The mixture was stirred for 2 hours and washed withwater (500 ml×2), 10% NaHCO₃ (aq.) (500 ml×2), water (500 ml×2) anddried over Na₂SO₄. The crude product was recrystalized from ethylacetate to give the title compound 2a as a white solid (23 g, 69%yield).

Synthesis of 3a (W₁=5-Cl, R₁=4-pyridyl)

To a solution of the 4-chloro-1′,1′,1′-trimethylacetanilide (a (509 mg,2.4 mmol) in THF (10 ml) under an atmosphere of nitrogen cooled to −78°C. was added n-butyllithium (2.5 ml, 2.4 M solution in diethyl ether, 6mmol) over 5 min. The reaction was left to warm up to 0° C. and kept atthis temperature for 2 hrs. Pyridine-4-carboxaldehyde (515 mg, 4.8 mmol)dissolved in THF (2 ml) was added and the reaction was left to slowlywarm to room temperature over 1 hr. The reaction was quenched with 1MHCl (10 ml) and then extracted with ethyl acetate (3×50 ml). Thecombined organic phase was dried over sodium sulfate, filtered and thesolvent removed in vacuum. The oily residue was passed through a plug ofsilica gel (5 g) using ethyl acetate as eluent. The material wasrecrystallized from ethyl acetate to yield compound 3a as white crystals(585 mg, 76% yield). clogP=3.23; R_(f) (petroleum ether:ethyl acetate(1:1)=0.31; HPLC (214 nm) t_(R)=8.20 (98.12%) min; ¹H NMR (400 MHz,CDCl₃) δ 1.04 (s, 9H), 5.74 (s, 1H), 7.07 (d, J=2.4 Hz, 1H), 7.24-7.30(m, 4H), 8.10 (d, J=8.8 Hz, 1H), 8.34 (d, J=5.7 Hz, 2H), 9.08 (s, 1H);¹³C NMR (100 MHz, CDCl₃) δ 27.2, 39.5, 73.9, 121.4, 124.4, 128.8, 129.0,132.5, 135.9, 149.0, 151.2, 177.1; ESMS m/z 319.3 [M+H]⁺, 637.3 [2M+H]⁺;LC/MS t_(R)=5.22 (319.1 [M+H]⁺, 637.1 [2M+H]⁺) min.

Synthesis of 10a (W₁=5-Cl, R₁=4-pyridyl)

A solution of the amide 3a (266 mg, 0.83 mmol) was dissolved in 3 M HClsolution (9 ml) and was heated at reflux for 4.5 hrs. After this timethe reaction was diluted with water (20 ml) and brought to basic pHusing 10M NaOH (4 ml). The aqueous solution was extracted withdichloromethane (3×50 ml). The combined organic phase was dried oversodium sulfate, filtered and the solvent removed in vacuum to yieldcompound 10a as an off white solid (189 mg, 96% yield) which wasanalytically pure and was used without further characterization.clogP=1.31; R_(f) (ethyl acetate)=0.36; ESMS m/z 235.1 [M+H]⁺; LC/MSt_(R)=3.90 (235.0 [M+H]⁺, 469.2 [2M+H]⁺) min.

Using similar procedures as outlined in 2 and 3, the followingbenzhydrol derivatives were prepared:

10b (W₁=5-Cl, R₁=2,6-dimethylphenyl) was synthesized from 2a and2,6-methylbenzaldehyde. Compound 10b was obtained as thick oil (285 mg,44% yield). clogP=5.54; R_(f) (petroleum ether:ethyl acetate (1:1)=0.41;HPLC (214 nm) t_(R)=9.42 (84.35%) min; ¹H NMR (400 MHz, CDCl₃) δ 1.21(s, 9H), 2.18 (s, 6H), 6.23 (s, 1H), 6.51 (d, J=2.4 Hz, 1H), 6.99-7.01(m, 2H), 7.11 (s, 1H), 7.13 (dd, J=2.4, 8.8 Hz, 1H), 8.04 (d, J=8.8 Hz,2H), 9.67 (s, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 20.9, 27.5, 39.8, 71.0,123.4, 127.0, 128.0, 128.2, 128.4, 128.5, 129.6, 130.9, 136.1, 136.6,137.1, 177.4; ESMS m/z 328.4 [M−OH]⁺, 346.5 [M+H]⁺; LC/MS t_(R)=9.74(328.1 [M−OH]⁺, 346.3 [M+H]⁺, 691.4 [2M+H]⁺) min.

10c (W₁=5-Cl, R₁=3-methyl-2-thiophenyl) was synthesized from 2a and3-methyl-2-thiophene carboxaldehyde. Compound 10c was used immediatelyin the subsequent step.

EXAMPLE 2

Synthesis of 5a (W₁=5-Cl, R₂=NH₂)

To a solution of N,O-dimethylhydroxylamine hydrochloride (2.0 g, 20mmol) dissolved in 90% ethanol (40 ml) was added triethylamine (2.5 ml,20 mmol). The reaction was left stirring for 10 min and 5-chloroisatoicanhydride (3 g, 15.2 mmol) was added to the above solution in smallportions. The reaction was heated to reflux for 1 h, after which time itwas poured into a 1:1 ice/saturated sodium bicarbonate solution (50 ml).The ethanol was removed under reduced pressure and the resulting aqueoussolution was extracted with ethyl acetate (3×50 ml). The combinedorganic phase was dried over magnesium sulfate, filtered and the solventremoved in vacuum to yield a colorless oil. This material was furtherpurified by flash chromatography on silica gel (150 g) using petroleumether:ethyl acetate 2:1 then 1:1 as eluent, to provide the desiredcompound 5a as a slightly off white solid (2.61 g, 70% yield)); R_(f)(petroleum ether:ethyl acetate (2:1)=0.42; HPLC (214 nm) t_(R)=5.50(94.6%) min; ¹H NMR (400 MHz, CDCl₃) δ 3.34 (s, 3H), 3.59 (s, 3H), 4.66(brs, 2H), 6.30 (dd, J=3.3, 8.6 Hz, 1H), 7.13 (dd, J=2.5, 8.6 Hz, 1H),7.36 (d, J=2.5 Hz, 1H); LC/MS t_(R)=5.44 (215.1 [M+H]⁺, 429.3 [2M+H]⁺,643.4 [3M+H]⁺) min.

In the case of starting with benzoic acid derivatives, thetransformation of 4 to 5 was carried out in the presence of EDC andDIEA.

Synthesis of 6a (W₁=5-Cl, R₁=3,5-dimethylphenyl, R₂=NH₂)

To a solution of 3,5-dimethyliodobenzene (1.59 g, 6.85 mmol) dissolvedin THF (10 ml) under an atmosphere of nitrogen cooled to −78° C. wasadded n-butyllithium (2.76 ml, 2.4 M solution in diethyl ether, 1equivalent based on the arylhalide). The reaction was left for 20 minafter which time the amide 5a (350 mg) dissolved in tetrahydrofuran (2.5ml) was added dropwise. The reaction was left stirring for a further 20min. The reaction was then quenched with the addition of 1M hydrochloricacid solution (5 ml) warmed to room temperature and diluted with ethylacetate (80 ml). The layers separated and the organic phase was washedwith water (1×20 ml), brine (1×20 ml), dried over sodium sulfate,filtered and the solvent removed in vacuum. The material was frozen in1:1 water/acetonitrile mixture and lyophilized to remove any volatilematerial. Compound 6a was obtained as a brown crystalline solid (450mg), which was analytically pure and used in the next step withoutfurther purification. clogP=3.25; R_(f) (petroleum ether:ethyl acetate(5:1)=0.66; ¹H NMR (400 MHz, CDCl₃) δ 2.37 (s, 6H), 6.00 (brs, 2H), 6.68(d, J=8.8 Hz, 1H), 7.21-7.26 (m, 4H), 7.41 (d, J=2.4 Hz, 1H); ESMS m/z260.0 [M+H]⁺; LC/MS t_(R)=10.83 (259.9 [M+H]⁺) min.

Synthesis of 6ad (W₁=5-CH₃, R₁=2-methylphenyl, R₂=OH)

Compound 6ac (W₁=5-CH₃, R₁=2-methylphenyl, R₂=—OCH₃) (0.094 g, 0.42mmol) was dissolved in anhydrous CH₂Cl₂ (3 ml) and cooled to 0° C. Borontribromide (0.180 ml, 1.87 mmol, 4.5 eq.) was added to cool solutiondropwise. The color of the solution changed from medium yellow to darkyellow-brown. The reaction was stirred at 0° C. for 1 hr. The progressof the reaction was monitored by TLC (ethyl acetate:petroleum ether=1:14). Water was added and the reaction solution partitioned betweenwater and CH₂Cl₂. The aqueous layer was back-extracted with CH₂Cl₂, thenthe combined organic fractions washed with brine, dried (Na₂SO₄) thenevaporated under reduced pressure to give a dark orange oil (0.11 g),which was purified using flash chromatography on silica (5 g) using 1:20diethyl ether:petroleum ether to give the target compound 6ad (0.06 g).R_(f) (1:5 diethyl ether:petrol)=0.56. HPLC (214 nm) t_(R)=10.03 min(96% overloaded).

Synthesis of 10d (W₁=5-Cl, R₁=3,5-dimethylphenyl, R₂=NH₂)

To the starting ketone 6a (250 mg, 0.96 mmol) dissolved in THF (5 ml)under an atmosphere of nitrogen cooled to 0° C. was added lithiumaluminum hydride (0.5 ml, 1.0M solution in diethyl ether). Analysis byTLC indicated the reaction was complete, saturated sodium bicarbonate(20 ml) was carefully added and the resultant solution was extractedwith ethyl acetate (3×50 ml). The combined organic phase was dried oversodium sulfate, filtered and the solvent removed in vacuum to yield 10das a brown oil which was used immediately in the next step.

Using similar procedures as outlined above, the following benzhydrolderivatives are prepared:

-   -   10e (W₁=5-Cl, R₁=1-methyl-2-imidazolyl, R₂=NH₂).    -   10f (W₁=5-Cl, R₁=2-benzothiazolyl, R₂=NH₂).    -   10g (W₁=5-Cl, R₁=2-thiophenyl, R₂=NH₂).    -   10h (W₁=5-Cl, R₁=1-methyl-2-pyrrolyl, R₂=NH₂).    -   10i (W₁=5-Cl, R₁=3-methylphenyl, R₂=NH₂).    -   10j (W₁=5-Cl, R₁=4-methylphenyl, R₂=NH₂).    -   10k (W₁=5-Cl, R₁=2,3-dimethylphenyl, R₂=NH₂).    -   10l (W=5-Cl, R₁=3,4-dimethylphenyl, R₂=NH₂).    -   10m (W₁=5-Cl, R₁=2,5-dimethylphenyl, R₂=NH₂).    -   10n (W₁=3,5-dichloro, R₁=2-methylphenyl, R₂=NH₂).    -   10o (W₁=3,5-dibromo, R₁=2-methylphenyl, R₂=NH₂).    -   10u (W₁=5-Cl, R₁=2-thiazolyl, R₂=NH₂)    -   10v (W₁=H, R₁=2,4-dimethylphenyl, R₂=NH₂)    -   10w (W₁=5-Cl, R₁=2,4-dimethylphenyl, R₂=NH₂)    -   10x (W₁=5-Cl, R₁=phenyl, R₂=NH₂)    -   10y (W₁=6-CH₃, R₁=2-amino-3-pyridinyl, R₂=NH₂)    -   10ab (W₁=5-CH₃, R₁=2-methylphenyl, R₂=NH₂)    -   10ac (W₁=H, R₁=2-methylphenyl, R₂=methoxy)    -   10ad (W₁=5-CH₃, R₁=2-methylphenyl, R₂=OH)

Synthesis of 10z (W₁=5-Cl, R₁=t-butyl, R₂=NH₂)

Step 1: To a solution of 4-bromo-3,5-dimethylpyrazole (700 mg, 0.93mmol) in THF (10 ml) under an atmosphere of nitrogen cooled to −78° C.was added t-butyllithium (10.6 ml, 1.3M solution in pentane, 13.8 mmol).The reaction was left for 15 min then2-amino-5-chloro-N-methoxy-N-methyl-benzamide (200 mg, 0.93 mmol)dissolved in THF (2 ml) was added. The reaction was left for 1 hr; thenit was quenched with 1M HCl (10 ml). The solution was diluted with brine(50 ml) and was extracted with ethyl acetate (3×50 ml). The combinedorganic phase was dried over sodium sulfate, filtered and the solventremoved in vacuum. The isolated product was the t-butyl ketone 6z(W₁=5-Cl, R₁=t-butyl, R₂=NH₂) rather than the desired pyrazolederivative. The residue was purified on silica gel (100 g) usingpetroleum spirit/ethyl acetate 10:1 as eluent. t-Butyl ketone 6z wasisolated as a yellow solid (175.5 mg, 89% yield). clogP=2.64; HPLC (214nm) t_(R)=7.78 (99%) min; ¹H NMR (400 MHz, CDCl₃) δ 1.37 (s, 9H), 5.32(brs, 2H), 6.62 (dd, J=3.2, 8.7 Hz, 1H), 7.14 (dd, J=2.4, 8.7 Hz, 1H),7.70 (d, J=2.4 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 28.6, 44.8, 119.0,119.6, 119.8, 129.5, 132.3, 147.8, 208.7; [M+H]⁺; LC/MS t_(R)=8.77(212.2 [M+H]⁺) min.

Step 2: To ketone 6z (250 mg, 0.96 mmol) dissolved in THF (5 ml) underan atmosphere of nitrogen cooled to 0° C. was added lithium aluminumhydride (0.5 ml, 1.0M solution in diethyl ether). Analysis by TLCindicated the reaction was complete, saturated sodium bicarbonate (20ml) was carefully added and the resultant solution was extracted withethyl acetate (3×50 ml). The combined organic phase was dried oversodium sulfate, filtered and the solvent removed in vacuum to yield 10z(W₁=5-Cl, R₁=t-butyl, R₂=NH₂) as a brown oil which was used immediatelyin the next step.

EXAMPLE 3

Synthesis of 8a (W₁=5-methyl)

A mixture of 2-amino-5-methylbenzoic acid (La) (2.0 g, 0.013 mol) and ARgrade THF (21 ml) was cooled to 0° C. With vigorous stirring, anhydroussodium carbonate (2.2 g, 0.021 mol, 1.6 eq.) was added followed bybenzoyl chloride (3.02 ml, 0.026 mol, 2 eq.) dropwise. The mixture wasleft to stand at 0° C. for an additional 30 min, after which the coldbath was removed and the mixture stirred at rt overnight. Water (15 ml)was added. The reaction mixture was evaporated under reduced pressure(to remove the THF), extracted with CH₂Cl₂ (×2). The combined CH₂Cl₂fractions were dried (Na₂SO₄) and evaporated under reduced pressure togive 8a as a cream colored solid (2.55 g, 82% yield). LC/MS t_(R)=8.16(256.1, acyclic amide), 9.47 (237.9 [M+H]⁺) min. The crude material wasused in the next step.

Synthesis of 9a (W₁=5-methyl, R₁=2-methylphenyl)

A solution of the crude material 8a from the previous step (0.5 g, 2mmol) in CH₂Cl₂ (15 ml) was cooled to −78° C. o-Tolyl magnesium bromide(2 M in diethyl ether, 2.3 ml, 5 mmol, 2.5 eq.) was added dropwise. Thereaction was monitored by TLC and was allowed to proceed at −78° C. fora total of 55 min. The reaction was briefly warmed to rt, whencesaturated NH₄Cl solution (2 ml) was added. The reaction mixture was thenpartitioned with CH₂Cl₂ (×2). The combined CH₂Cl₂ fractions wereback-extracted with brine (×2), then evaporated under reduced pressureto give crude 9a (W₁=5-methyl, 2-methylphenyl) as a yellow oil (0.58 g).LC/MS t_(R)=5.59 (no identifiable ion); 8.41 (195.1, unidentified); 9.45(237.9, (8a)); 10.73 (329.9 (9a)) min. R_(f) (ethyl acetate:petroleum40-60 (1:20))=0.19 (target compound), 0.27 (starting material (a)).Silica gel column chromatography was attempted using this solventsystem, however co-elution of the two bands was observed. The crudematerial was therefore taken to the next step. The yield of productafter purification by chromatography was 0.44 g.

Synthesis of 6b (W₁=5-methyl, R₁=2-methylphenyl, R₂=NH₂)

The crude product 9a from the above reaction (0.44 g), MeOH (5 ml), H₂O(4 ml) and sodium hydroxide pellets (3.2 g, 0.08 mol, 100 eq.) werestirred and heated to reflux for 4 hrs, then cooled to rt and water (30ml) added. The mixture was partitioned with CH₂Cl₂ (×4) and the combinedCH₂Cl₂ fractions back-extracted with brine (×2). The combined CH₂Cl₂fractions were then dried (Na₂SO₄) and evaporated under reduced pressureto give 6b as a dark yellow oil which was dried under vacuum overnight(0.153 g). LC/MS t_(R) _(=8.34) (195.0, unidentified), 8.60 (226.1,[M+H]⁺), 8.79 (no identifiable ion) min.

Synthesis of 10p (W₁=5-methyl, R₁=2-methylphenyl, R₂=NH₂)

The crude reaction product of 6b from above step (0.13 g) was dissolvedin THF (5 ml) then cooled to ice/water temperature. Lithium aluminumhydride (nominal I M solution in diethyl ether) was added (1 ml) under astream of nitrogen via syringe. The reaction was monitored using TLC.Additional aliquots of 0.5 ml of LiAlH₄/Et₂O solution were added to thereaction solution at the 35 min and 2 hr 10 min marks. The reaction wasallowed to proceed for 3 hr in total. Saturated NaHCO₃ solution wasadded. The mixture was partitioned between EtOAc and water. Afterseveral extractions, the combined EtOAc fractions were dried (Na₂SO₄)then evaporated under reduced pressure to yield 10p as a yellow oil. Thecrude reaction product was carried through to the next step.

Using similar procedures as outlined above, the following benzhydrolderivatives are prepared:

-   -   10q (W₁=H, R₁=2-methylphenyl, R₂=NH₂)    -   10r (W₁=H, R₁=2-chlorophenyl, R₂=NH₂)    -   10x (W₁=5-Cl, R₁=phenyl, R₂=NH₂)    -   10aa (W₁=5-Cl, R₁=2-methylphenyl, R₂=NH₂):

Using commercially available benzophenones, the following benzhydrolderivatives were prepared using the procedure outlined in the synthesisof 10d.

-   -   10s (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂).    -   10t (W₁=5-nitrol, R₂=phenyl, R₂=NH₂).    -   10ae (W₁=H, R₁=5-amino-2-chlorophenyl, R₂=NH₂) using LiBH₄ in        chlorophenyl THF.    -   10af (W₁=H, R₁=3-amino-4-chlorophenyl, R₂=NH₂) was obtained from        4-chloro-3-nitrobenzophenone by reduction using activated iron        in acetic acid/water at 90° C., followed by reduction using        LiBH₄ in THF.    -   10ag (W₁=5-Cl, R₁=2-chlorophenyl, R₂=OH).

p-Chlorophenol (2 g, 15 mmol), methyl iodide (1.36 ml, 21.8 mmol) andpotassium carbonate (2.07 g, 15.6 mmol) in dry acetone (15 ml) weretaken in a 50 ml flask. After refluxing for 3 hrs at 60° C., acetone wasremoved using a rotary evaporator. Water was added and the product wasextracted with CH₂Cl₂ to afford 2.2 g of crude product, which waspurified by column chromatography using hexane as eluent to yieldp-chloroanisole in 75% yield (1.59 g).

p-Chloroanisole (400 mg, 2.8 mmol) and AlCl₃ (410 mg, 3 mmol) in dry CS₂(10 ml) were taken in 2-neck 25 ml rb flask. After refluxing for 15 minat 50° C. 2-chloro benzoyl chloride (420 μL, 3.36 mmol) was added viasyringe and the mixture was refluxed for 4 hrs. The reaction wasquenched by adding 5 ml of 1M HCl, extracted with DCM, washed withwater, dried over Na₂SO₄ and concentrated to afford 1 g of the crudeproduct. The product was purified by column chromatography using 20%CHCl₃/pet ether to yield 700 mg (72%) of compound 6ag (W₁=5-Cl,R₁=2-chlorophenyl, R₂=OCH₃). m.p. 104.5-108.1° C. ¹H NMR (300 MHz,CDCl₃) δ 11.88 (s, 1H), 7.52-7.33 (m, 5H), 7.2 (d, J=2.7 Hz, 1H), 7.03(d, J=9 Hz, 1H).

Compound 6ag (100 mg, 0.3 mmol) in dry THF (2 ml) was taken in a 10 mlrb flask and cooled to 0° C. To the cold stirred solution LAH (28 mg,0.6 mmol) was added and stirred for 8 hrs. A saturated solution ofsodium potassium tartarate (5 ml) was added into the reaction mixtureand stirring was continued for 30 min. After the separation of organicand aqueous layers, the reaction mixture was extracted with DCM, washedwith water and dried over Na₂SO₄ to afford 78 mg (75%) of compound 10ag.

-   -   10ah (W₁=5-Cl, R₁=phenyl, R₂=—OCH₃).

5-Chloro-2-hydroxybenzophenone (500 mg, 2.5 mmol), methyl iodide (0.22ml, 3.5 mmol) and potassium carbonate (0.37 g, 2.7 mmol) in dry acetone(15 ml) were taken in a 25 ml flask. After refluxing for 3 hrs, acetonewas removed on a rotavap, water was added and the product was extractedwith CH₂Cl₂ to afford 0.620 g of compound 4. The product was purified bycolumn chromatography hexane as eluent to yield 0.413 g (78%) of5-chloro-2-methoxybenzophenone.

5-chloro-2-methoxybenzophenone (200 mg, 0.94 mmol) in dry THF (5 ml) wascooled to 0° C. To the cold stirred solution LAH (115 mg, 3 mmol) wasadded and stirring was continued for 8 hrs. A saturated solution ofsodium potassium tartarate (5 ml) was added into the reaction mixtureand stirring was continued for 30 min. After the separation of organicand aqueous layers, the reaction mixture was extracted with DCM, washedwith water and dried over Na₂SO₄ and concentrated to afford 168 mg (83%)of analytically pure 1_ah.

Synthesis of 10ai (W₁=5-Cl, R₁=phenyl, R₂=OH)

To a solution of 5-chloro-2-hydroxybenzophenone (100 mg, 0.429 mmol) indry THF (0.5 ml) at 0° C. under argon atmosphere was added LAH (35 mg,0.92 mmol). The mixture was stirred for 8 hrs. The reaction mixture wasadded to 6 M HCl (2 ml) 0° C., neutralized with satd. NaHCO₃. Theproduct was extracted with DCM (3×10 ml) dried over anhydrous Na₂SO₄ andconcentrated to afford analytically pure 10ai (2 g, 78%).

Synthesis of 10aj (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂)

To a solution of pentyl nitrite (2.5 mL, 19.2 mmol) in THF (25 mL) at65° C. was added a solution of compound2-amino-2′,5-dichlorobenzophenone (2 g, 7.5 mmol) in THF (10 mL) over aperiod of 1 hr. The reaction mixture was refluxed for 3 hrs and THF wasremoved on a rotary evaporator. The residue was extracted with benzene,washed with 16% H₂SO₄ (20 mL), dried over anhyd. Na₂SO₄ and concentratedto afford 3.6 g of the crude product which was purified by columnchromatography using 5% ethyl acetate/petroleum ether to yield 6aj(W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂) (1.8 g, 62%). ¹H NMR (300 MHz,CDCl₃) δ 7.79 (s, 1H), 7.66 (d, J=7.8 Hz, 1H), 7.56 (dd, J=8.1 Hz, J=1.2Hz, 1H), 7.48-7.38 (m, 5H).

To a solution of compound 6ai (30 mg, 0.12 mmol) in THF (0.2 mL) wasadded NaBH₄ (12 mg, 0.32 mmol) in THF (0.4 mL) followed by 1 mL ofwater. The reaction mixture was allowed to stir at room temperature for48 hrs. THF was removed on a rotary evaporator, the residue wasextracted with chloroform, washed with water and dried over anhyd.Na₂SO₄ to afford 1a (16 mg, 52%), which was used in the next step without further purification.

EXAMPLE 4

10ak, 10al, 10 am and 10an were prepared using a slightly modifiedversion of the procedure set forth in Haider et al., J. HeterocyclicChem., 27:1645 (1990) and Ubeda et al., Synthesis, 1176 (1998).

EXAMPLE 5

Synthesis of Thioether Ester 11a (W₁=5-Cl, R₁=4-pyridyl, R₂=NH₂)

A solution of 10a in methyl thioglycolate (0.5 ml) and TFA (2 ml) wasstirred at room temperature for 18 hrs. After this time the solvent wasremoved in vacuum and the residue was diluted with dichloromethane (70ml) and washed with 1 M NaOH (20 ml) and then with brine (20 ml). Thecombined aqueous phases were back extracted with dichloromethane (20ml). The combined organic phase was dried over sodium sulfate, filteredand the solvent removed in vacuum. An attempt at purification by flashchromatography-on silica gel (25 g) using ethyl acetate, then ethylacetate/methanol 9:1 as eluent, failed to give a pure sample of thedesired compound. Preparative HPLC conditions seemed to decompose mostof the compound, only a small sample was obtained from the HPLCpurification. 11a: brown oil (4.1 mg, 3% yield); clogP=2.20; R_(f)(ethyl acetate)=0.49; HPLC (214 nm) t_(R)=5.98 (76.80%) min; ¹H NMR (400MHz, CDCl₃) δ 3.24 (d, J=16.3 Hz, 1H), 3.33 (d, J=16.3 Hz, 1H), 3.65 (s,1H), 5.53 (s, 1H), 6.77 (d, J=8.6 Hz, 1H), 6.80 (d, J=2.4 Hz, 1H), 7.09(dd, J=2.4, 8.4 Hz, 1H), 8.00 (d, J=5.0 Hz, 2H), 8.71 (d, J=5.0 Hz, 2H);ESMS m/z 323.3 [M+H]⁺; LC/MS t_(R)=5.68 (323.0 [M+H]⁺, 645.0 [2M+H]⁺)min.

Synthesis of Thioether Ester 11b (W₁=5-Cl, R₁=2,6-dimethylphenyl,R₂=NH₂) and 11ba (W₁=5-Cl, R₁=2,6-dimethylphenyl, R₂=—NHC(═O)CF₃)

A solution of the amine 10b from above in methyl thioglycolate (0.5 ml)and TFA (2 ml) was stirred at room temperature for 18 hrs. After thistime analysis by LCMS showed a mixture of the desired product plusstarting material. The reaction was then heated to 60° C. for 15 hrs.The solvent was removed under a stream of nitrogen. The residue wasdiluted with dichloromethane (80 ml) and wash with brine (1×20 ml), thenwith 1M NaOH (1×40 ml) and then finally with brine (1×20 ml). Thecombined organic phase was dried over sodium sulfate, filtered and thesolvent removed in vacuum. Analysis by LCMS showed a mixture of twocompounds. The two compounds were separated by flash chromatography onsilica gel (50 g) using petroleum spirit/ethyl acetate, 10:1 then 5:1 aseluent. The first compound to elute off the column was trifluoroanilideof 11ba, isolated as a slightly colored oil (46.5 mg, 20% yield);clogP=5.37; R_(f) (petroleum spirit/ethyl acetate, 5:1)=0.60; HPLC (214nm) t_(R)=9.53 (97%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.26 (s, 6H), 3.19(d, J=15.5 Hz, 1H), 3.27 (d, J=15.5 Hz, 1H), 3.71 (s, 3H), 5.85 (s, 1H),7.08 (d, J=7.5 Hz, 2H), 7.17 (apparent t, J=7.5 Hz, 1H), 7.34 (dd,J=2.3, 8.6 Hz, 1H), 7.58 (d, J=2.3 Hz, 1H), 7.90 (d, J=8.6 Hz, 1H), 8.56(s, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 21.4, 33.1, 45.9, 52.6, 118.5 (q,J=288.9 Hz), 125.4, 128.4, 128.7, 129.9, 130.3, 131.5, 132.1, 132.2,132.6, 137.9, 155.1 (q, J=37.4 Hz), 170.4; LC/MS t_(R)=9.81 (340.0[M—C₃H₅O₂S]⁺, 462.9 [M+H₂O]⁺, 890.9 [2M+H]⁺, 908.4 [2M+H₂O)⁺) min.

The second compound to elute off the column was 11b, isolated as aslightly colored oil (28.2 mg, 15% yield); clogP=4.51; R_(f) (petroleumspirit/ethyl acetate, 5:1)=0.37; HPLC (214 nm) t_(R)=9.39 (94%) min; ¹HNMR (400 MHz, CDCl₃) δ 2.33 (s, 6H), 3.16 (d, J=15.5 Hz, 1H), 3.25 (d,J=15.5 Hz, 1H), 3.71 (s, 3H), 4.00 (brs, 2H), 5.64 (s, 1H), 6.59 (d,J=8.4 Hz, 1H), 7.02-7.06 (m, 3H), 7.11 (m, 1H), 7.30 (d, J=1.8 Hz, 1H);¹³C NMR (100 MHz, CDCl₃) δ 21.7, 33.4, 46.5, 52.5, 117.8, 123.0, 123.8,127.9, 128.0, 129.3, 129.9, 133.9, 138.0, 144.0, 171.1; LC/MS t_(R)=9.79(244.1 [M−C₃H₅O₂S]⁺, 350.0 [M+H]⁺, 701.0 [2M+H]⁺) min.

Synthesis of Thioether Ester 11c (W₁=5-Cl, R₁, =3-methyl-2-pyrrolyl,R₂=NH₂)

Alcohol 10c from above was dissolved in dichloromethane (5 ml), methylthioglycolate (50 μL) was added followed by TFA (50 μL). After 15 min,TLC indicated the consumption of starting material along with theformation of a number of other products. The solvent was removed invacuum and the residue was purified on silica gel (50 g) using gradientelution starting with petroleum spirit/ethyl acetate 10:1 to 2:1. Thethird major fraction off the column was found to be the desired compound11c, isolated as brown oil (18.5 mg, 12.2% yield); clogP=4.01; R_(f)(petroleum spirit/ethyl acetate, 4:1)=0.21; HPLC (214 nm) t_(R)=11.41(85.89%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.01 (s, 3H), 3.28 (d, J=16.5Hz, 1H), 3.37 (d, J=16.5 Hz, 1H), 3.69 (brs, 2H), 3.75 (s, 3H), 5.56 (s,1H), 6.65 (d, J=8.4 Hz, 1H), 6.87 (d, J=5.1 Hz, 1H), 6.96 (d, J=2.4 Hz,1H), 7.03 (dd, J=2.4, 8.4 Hz, 1H), 7.21 (d, J=5.1 Hz, 1H); ¹³C NMR (100MHz, CDCl₃) δ 13.7, 33.9, 42.8, 52.6, 117.7, 123.4, 124.1, 125.6, 128.2,128.6, 131.0, 135.8, 136.9, 143.2, 171.5; ESMS In/z 341.9 [M+H]⁺, 383.2[M+NH₄]⁺; LC/MS t_(R)=9.24 (342.1 [M+H]⁺, 683.2 [2M+H]⁺) min.

Synthesis of Thioether Ester of 11d (W₁=5-Cl, R₁=3,5-dimethylphenyl,R₂=NH₂)

(i) To the alcohol 10d from above was added methyl thioglycolate (500μL, 5.6 mmol) followed by TFA (2 ml). The reaction was heated at 85° C.overnight. The next morning, the black solution was diluted withdichloromethane (80 ml) and washed with 1M sodium hydroxide (20 ml)followed by brine (20 ml). The organic phase was dried over sodiumsulfate, filtered and the solvent removed in vacuum to yield a creamyamorphous solid (300 mg). Analysis of the solid by LCMS indicated amixture of three compounds whose analytical data was consistent with thebenzothiazepine, the thioether methyl ester 11d, as well as thethioether carboxylic acid (this acid was probably formed during theworkup procedure whereby some of the methyl ester 11d was hydrolyzed bythe basic wash solution). The mixture was treated with WSC, step (ii)below, in order to convert the acid to the cyclized benzothiazepine andsimplify the purification of the reaction mixture.

(ii) To a solution of the mixture above in tetrahydrofuran (50 ml) wasadded diisopropylethylamine (125uL, 0.72 mmol), followed by EDC(WSC.HCl) (136 mg, 0.71 mmol) and finally dimethylaminopyridine (10.6mg, 0.09 mmol). The reaction was left to stir overnight at roomtemperature. Next morning TLC indicated complete consumption of thecarboxylic acid resulting in a mixture of two products; assumed to bethe desired thio ether methyl ester 11d, as well as the cyclizedbenzothiazepine. The solvent was removed in vacuum and the residues wastaken up into ethyl acetate (70 ml) and washed with 10% citric acid(1×30 ml), saturated sodium bicarbonate (1×30 ml) and finally with brine(1×30 ml). The organic phase was dried over magnesium sulfate, filteredand the solvent removed in vacuum. The residue was purified by flashchromatography on silica gel (25 g) using petroleum ether:ethyl acetate2:1 as eluent. The first compound to elute from the column was 11disolated as an off white solid (41.4 mg, 12.1% yield over three steps);clogP=4.51; R_(f) (petroleum ether:ethyl acetate (2:1)=0.57; HPLC (214nm) t_(R)=9.65 (88.0%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.32 (s, 6H), 3.13(d, J=16.0 Hz, 1H), 3.18 (d, J=16.0 Hz, 1H), 3.72 (s, 3H), 6.62 (d,J=8.4 Hz, 1H), 6.93 (s, 1H), 6.96 (d, J=2.5 Hz, 1H), 7.02 (dd, J=2.5,8.4 Hz, 1H), 7.07 (s, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 21.32, 33.04,49.7, 52.5, 117.6, 123.0, 126.1, 126.8, 128.2, 128.9, 129.6, 137.7,138.4, 143.6, 171.2; ESMS m/z 244.0 [M−C₃H₄O₂S]⁺, 350.2 [M+H]⁺; LC/MSt_(R)=9.89 (244.0 [M−C₃H₄O₂S]⁺, 350.1 [M+H]⁺, 699.3 [2M+H]⁺) min.

Synthesis of Thioether Ester 11e (W₁=5-Cl, R₁=-methyl-2-imidazolyl,R₂=NH₂)

A sample of 10e (50 mg, 0.21 mmol) from above was mixed with methylthioglycolate (500 μL) followed by TFA (2 ml). The reaction was left tostir at room temperature for 24 h. After this time the reaction wasdiluted with dichloromethane (100 ml) and was washed once with a 1:1mixture of brine/1M NaOH (50 ml). The aqueous phase was extracted withdichloromethane (2×20 ml). The combined organic phase was washed withbrine (1×50 ml), dried over sodium sulfate, filtered and the solventremoved in vacuum to yield a greenish solid (55 mg). Analysis of thereaction showed a very messy reaction but the main peak by LCMS had a MSconsistent with the desired product. The compound was dissolved indichloromethane (1 ml) and was carefully loaded onto two preparative TLCplates. The plates were developed using a mixture consisting of 40 mlethyl acetate and 2 ml concentrated ammonia solution. The main UV activeband was cut from the plate and the desired product was eluted from theplate by washing with ethyl acetate (250 ml). The solvent was removed invacuum to yield a white solid (>100 mg probably silica gel). The solidwas taken up into chloroform (2 ml) and the crystalline solid wasremoved by filtration to yield 11e as a brown oil (20 mg, 29% yield);clogP=1.73; R_(f) (ethyl acetate/5% NH₄OH)=0.59-0.70); HPLC (214 nm)t_(R)=5.81 (75.92%) min; ¹H NMR (400 MHz, CDCl₃) δ 3.16 (d, J=15.8 Hz,1H), 3.28 (d, J=15.8 Hz, 1H), 3.48 (s, 3H), 3.72 (s, 3H), 4.50 (brs,2H), 5.50 (s, 1H), 6.89 (dd, J=2.0, 9.1 Hz, 1H), 6.83 (d, J=1.1 Hz, 1H),7.01 (d, J=1.1 Hz, 1H), 7.03-7.05 (m, 2H); ESMS m/z 326.1 [M+H]⁺; LC/MSt_(R)=5.21 (325.9 [M+H]⁺) min.

11f (W₁=5-Cl, R₁=2-benzothiazolyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, 11f was preparedand isolated as a yellow oil (21.9 mg, 20.6% yield); clogP=4.41; R_(f)(petroleum ether:ethyl acetate (2:1)=0.41; HPLC (214 nm) t_(R)=8.84(76.8%) min; ¹H NMR (400 MHz, CDCl₃) δ 3.32 (d, J=16.1 Hz, 1H), 3.42 (d,J=16.1 Hz, H), 3.72 (s, 3H), 5.80 (s, 1H), 6.64 (d, J=8.6 Hz, 1H), 7.07(dd, J=2.4, 8.6 Hz, 1H), 7.14 (d, J=2.4 Hz, 1H), 7.40 (m, 1H), 7.49 (m,1H), 7.87 (d, J=7.9 Hz, 1H), 8.02 (d, J=8.1 Hz, 1H); ESMS m/z 379.3[M+H]⁺; LC/MS t_(R)=9.11 (379.0 [M+H]⁺) min.

11g (W₁=5-Cl, R₁=2-pyrrolyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, 11g was preparedand isolated as a yellow oil (21.9 mg, 20.6% yield); clogP=4.41; R_(f)(petroleum ether:ethyl acetate (2:1)=0.41; HPLC (214 nm) t_(R)=8.84(76.8%) min; ¹H NMR (400 MHz, CDCl₃) δ 3.32 (d, J=16.1 Hz, 1H), 3.42 (d,J=16.1 Hz, H), 3.72 (s, 3H), 5.80 (s, 1H), 6.64 (d, J=8.6 Hz, 1H), 7.07(dd, J=2.4, 8.6 Hz, 1H), 7.14 (d, J=2.4 Hz, 1H), 7.40 (m, 1H), 7.49 (m,1H), 7.87 (d, J=7.9 Hz, 1H), 8.02 (d, J=8.1 Hz, 1H); ESMS m/z 379.3[M+H]⁺; LC/MS t_(R)=9.11 (379.0 [M+H]⁺) min.

11h (W₁=5-Cl, R₁=1-methyl-2-pyrrolyl R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, compound 11h wasprepared and isolated as a creamy amorphous solid (55 mg); clogP=2.32;R_(f) (petroleum ether:ethyl acetate (2:1)=0.56; HPLC (214 nm)t_(R)=8.75 (96.47%) min; ¹H NMR (400 MHz, CDCl₃) δ 3.18 (d, J=16.3 Hz,1H), 3.32 (s, 3H), 3.53 (d, J=16.3 Hz, 1H), 3.70 (s, 3H), 4.32 (brs,1H), 5.32 (s, 1H), 6.09 (t, J=3.1 Hz, 1H), 6.36 (m, 1H), 6.59-6.61 (m,2H), 6.84 (d, J=2.5 Hz, 1H), 7.00 (dd, J=2.5, 8.6 Hz, 1H); ¹³C NMR (100MHz, CDCl₃) 629.6, 33.3, 33.6, 41.9, 52.5, 107.0, 109.8, 117.7, 123.3,123.6, 124.7, 128.5, 128.6, 143.5, 171.6; ESMS m/z 219.3 [M−C₃H₄O₂S]⁺,325.1 [M+H]⁺; LC/MS t_(R)=8.88 (325.0 [M+H]⁺, 649.3 [2M+H]⁺) min.

Synthesis of thioether ester 11i-m

(i) To the alcohol 10i from step 6 was added methyl thioglycolate (500μL, 5.6 mmol) followed by TFA (2 ml). The reaction was sealed with astopper and heated at 60° C. 16 hrs. The solution was diluted withdichloromethane (80 ml) and washed with 1 M sodium hydroxide (20 ml)followed by brine (20 ml). The organic phase was dried over sodiumsulfate, filtered and the solvent removed in vacuum to yield a creamyamorphous solid. Analysis of the solid by LMS indicated that only thethioether methyl ester was present along with a small amount of the acidformed by hydrolysis of the methyl ester during workup. The resultingreaction mixture above was diluted with 6 ml of methanol and 2 ml wasremoved for (ii) below.

(ii) To the methanol solution above (2 ml) was added 1 M sodiumhydroxide solution (1 ml). The reaction was left to stir at roomtemperature for 1 hr. After this time the reaction was diluted withbrine (30 ml) and neutralized with 10% HCl solution. The aqueous layerwas extracted with ethyl acetate (3×50 ml). The combined organic phasewas dried over sodium sulphate, filtered and the residue combined withthe remaining material from (i) above and used in (iii) below.

(iii) To a solution of the mixture above in tetrahydrofuran (20 ml) wasadded diisopropylethylamine (67 mL, 0.39 mmol), followed by EDC (74 mg,0.39 mmol) and dimethylaminopyridine (4.0 mg, 0.03 mmol). The reactionwas left to stir overnight at room temperature. TLC indicated completeconsumption of the carboxylic acid. The resulting mixture contained twoproducts: the desired thio ether methyl ester 11i and the cyclizedcorresponding benzothiazepine. The solvent was removed in vacuum and theresidues were taken up into ethyl acetate (70 ml) and washed with 10%citric acid (1×30 ml), saturated sodium bicarbonate (1×30 ml) andfinally with brine (1×30 ml). The organic phase was dried over sodiumsulfate, filtered and the solvent removed in vacuum. The residue waspurified by flash chromatography on silica gel (25 g) using petroleumether:ethyl acetate 2:1 as eluent. 11i (W₁=5-Cl, R₁=3-methylphenyl,R₂=NH₂): isolated as a brown oil (52.2 mg, 38.2% yield); clogP=4.03;R_(f) (petroleum ether:ethyl acetate (2:1)=0.65; HPLC (214 nm)t_(R)=8.80 (94.95) min; ¹H NMR (400 MHz, CDCl₃) δ 2.28 (s, 3H), 3.06 (d,J=16.1 Hz, 1H), 3.11 (d, J=16.1 Hz, 1H), 3.60 (brs, 2H), 3.64 (s, 3H),5.27 (s, 1H), 6.55 (d, J=8.4 Hz, 1H), 6.86 (d, J=2.4 Hz, 1H), ), 6.94(dd, J=2.4, 8.4 Hz, 1H), ), 7.04 (d, J=7.5 Hz, 1H), 7.17-7.24 (m, 3H);¹³C NMR (100 MHz, CDCl₃) δ 21.4, 33.3, 49.7, 52.5, 117.7, 123.1, 126.1,128.3, 128.7, 128.9, 129.7, 137.7, 138.5, 143.5, 171.2; ESMS m/z 230.3[M−C₃H₄O₂S]⁺, 336.3 [M+H]⁺]⁺, 671.2 [2M+H]⁺; LC/MS t_(R)=9.43 (229.9[M−C₃H₄O₂S]⁺, 336.1 [M+H]⁺, 671.1 [2M+H]⁺) min.

Using similar procedure as (i), (ii) and (iii) 11j-m were also prepared.

11j (W₁=5-Cl, R₁=4-methylphenyl, R₂=NH₂): isolated as a brown oil (40.0mg, 29.3% yield); clogP=4.03; R_(f) (petroleum ether:ethyl acetate(2:1)=0.65; HPLC (214 nm) t_(R)=8.90 (94.39) min; ¹H NMR (400 MHz,CDCl₃) δ 2.28 (s, 3H), 3.03 (d, J=16.0 Hz, 1H), 3.10 (d, J=16.0 Hz, 1H),3.64 (s, 2H), 4.23 (brs, 3H), 5.26 (s, 1H), 6.71 (d, J=8.5 Hz, 1H), 6.86(d, J=2.4 Hz, 1H), ), 6.94 (dd, J=2.4, 8.5 Hz, 1H), ), 7.11 (d, J=8.0Hz, 1H), 7.29 (d, J=8.0 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 21.1, 33.3,49.6, 52.5, 117.6, 123.0, 126.0, 128.3, 128.8, 128.9 (double intensity),129.5 (double intensity), 134.7, 137.6, 143.5, 171.2; ESMS m/z 230.3[M−C₃H₄O₂S]⁺, 336.3 [M+H]⁺, 671.2 [2M+H]⁺; LC/MS t_(R)=9.51 (230.3[M−C₃H₄O₂S]⁺, 336.1 [M+H]⁺, 671.1 [2M+H]⁺) min.

11k (W₁=5-Cl, R₁=2,3-dimethylphenyl, R₂=NH₂): isolated as a brown oil(60.0 mg, 44.5% yield); clogP=4.51; R_(f) (petroleum ether:ethyl acetate(2:1)=0.65; HPLC (214 nm) t_(R)=9.82 (88.81) min; ¹H NMR (400 MHz,CDCl₃) δ 1.97 (s, 3H), 2.20 (s, 3H), 3.05 (d, J=16.2 Hz, 1H), 3.11 (d,J=16.2 Hz, 1H), 3.63 (s, 3H), 4.05 (brs, 2H), 5.53 (s, 1H), 6.56 (d,J=8.4 Hz, 1H), 6.72 (d, J=2.4 Hz, 1H), 6.96 (dd, J=2.4, 8.4 Hz, 1H),7.05 (d, J=7.1 Hz, 1H), 7.10 (apparent t, J=7.5 Hz, 1H), 7.61 (d, J=7.5Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 14.4, 20.7, 33.4, 46.6, 52.4, 117.5,123.1, 125.7, 125.9, 126.2, 128.2, 128.5, 129.5, 135.4, 135.6, 137.5,143.5, 171.4; ESMS m/z 244.3 [M−C₃H₄O₂S]⁺, 350.2 [M+H]⁺; LC/MSt_(R)=9.88 (244.0 [M−C₃H₄O₂S]⁺, 350.0 [M+H]⁺, 699.1 [2M+H]⁺) min.

11l (W₁=5-Cl, R₁=3,4-dimethylphenyl, R₂=NH₂): isolated as a brown oil(56.0 mg, 41.6% yield); clogP=4.51; R_(f) (petroleum ether:ethyl acetate(2:1)=0.65; HPLC (214 nm) t_(R)=9.71 (85.37) min; ¹H NMR (400 MHz,CDCl₃) δ 2.18 (s, 6H), 3.04 (d, J=16.1 Hz, 1H), 3.09 (d, J=16.1 Hz, 1H),3.64 (s, 3H), 4.05 (brs, 2H), 5.23 (s, 1H), 6.53 (d, J=8.3 Hz, 1H), 6.89(d, J=2.4 Hz, 1H), 6.93 (dd, J=2.4, 8.3 Hz, 1H), 7.05 (d, J=7.5 Hz, 1H),7.12-7.17 (m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 19.4, 19.8, 33.4, 49.5,52.5, 117.6, 123.0, 126.1, 126.3, 128.2, 128.8, 130.0, 130.2, 135.1,136.3, 137.0, 143.5, 171.2; ESMS m/z 244.2 [M−C₃H₄O₂S]⁺, 350.3 [M+H]⁺;LC/MS t_(R)=9.76 (244.1 [M−C₃H₄O₂S]⁺, 350.0 [M+H]⁺, 699.2 [2M+H]⁺) min.

11m (W₁=5-Cl, R₁=2,5-dimethylphenyl, R₂=NH₂): isolated as a brown oil(51.0 mg, 37.9% yield); clogP=4.51; R_(f) (petroleum ether:ethyl acetate(2:1)=0.65; HPLC (214 nm) t_(R)=9.79 (76.77) min; ¹H NMR (400 MHz,CDCl₃) δ 2.04 (s, 3H), 2.30 (s, 3H), 3.06 (d, J=16.0 Hz, 1H), 3.11 (d,J=16.0 Hz, 1H), 3.63 (s, 3H), 4.05 (brs, 2H), 5.44 (s, 1H), 6.55 (d,J=8.4 Hz, 1H), 6.76 (d, J=2.2 Hz, 1H), 6.92-6.99 (m, 3H), 7.51 (s, 1H);¹³C NMR (100 MHz, CDCl₃) δ 18.5, 21.2, 33.4, 46.2, 52.4, 117.5, 123.1,125.6, 128.2, 128.4 (double intensity), 128.8, 130.8, 134.0, 135.3,135.8, 143.7, 171.4; ESMS m/z 244.2 [M−C₃H₄O₂S]⁺, 350.3 [M+H]⁺; LC/MSt_(R)=9.87 (244.0 [M−C₃H₄O₂S]⁺, 350.1 [M+H]⁺, 699.2 [2M+H]⁺) min.

11n (W₁=3,5-dichloro, R₁=2-methylphenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11n was preparedas a light yellow solid (9 mg). HPLC (214 nm) t_(R)=9.62 min. ¹H NMR(400 MHz, CDCl₃) δ 2.10 (s, 3H), 3.14 (dd, J=28.4, 16.4 Hz, 2H), 3.71(s, 3H), 4.86 (bs, 2H, NH₂), 5.54 (s, 1H), 6.66 (d, J=2.4 Hz, 1H),7.15-7.30 (m, 4H), 7.80 (d, J=7.2 Hz, 1H). LCMS t_(R)=10.67 min (369.9[M+H]⁺).

11o (W₁=3,5-dibromo, R₁=2-methylphenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11o was preparedas an orange-yellow oil which solidified upon standing (35 mg). HPLC(214 nm) t_(R)=9.97 min. ¹H NMR (400 MHz, CDCl₃) δ 2.11 (s, 3H), 3.13(dd, J=29.2, 16.4 Hz, 2H), 3.71 (s, 3H), 4.95 (bs, 2H, NH₂), 5.53 (s,1H), 6.82 (d, J=2.4 Hz, 1H), 7.15-7.30 (m, 3H), 7.45 (d, J=2.4 Hz, 1H),7.80 (d, J=7.2 Hz, 1H). LCMS t_(R)=10.96 min (459.9 [M+H]⁺).

11p (W₁=5-methyl, R₁=2-methylphenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11p was preparedand isolated as an off-white solid (0.046 g after freeze-drying). R_(f)(EtOAc:petrol (1:10))=0.13. HPLC (214 nm) t_(R)=7.82 (88.2%) min; ¹H NMR(400 MHz, CDCl₃) δ 2.11 (s, 3H), 2.19 (s, 3H), 3.15 (dd, J=24, 16 Hz,2H), 3.71 (s, 3H), 4.11 (bs, 2H), 5.61 (s, 1H), 6.62-6.66 (m, 2H), 6.87(dd, J=8, 1.6 Hz, 1H), 7.15-7.30 (m, 3H), 7.82 (d, J=7.6 Hz, 1H); LC/MSt_(R)=7.72 (316.1 [M+H]⁺) min.

11r (W₁=H, R₁=2-chlorophenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11r was obtainedas a yellow powder (360 mg, 1.11 mmol, 41% yield for three steps): R_(f)(petroleum ether:ethyl acetate (2:1)=0.60). HPLC (214 nm) t_(R)=8.51(80.0%) min. ¹H NMR (400 MHz, CDCl₃) δ 3.11 (d, J=16.0 Hz, 1H), 3.18 (d,J=16.0 Hz, 1H), 3.68 (s, 3H), 5.82 (s, 1H), 6.63 (ddd, J=7.6, 7.6, 0.8Hz, 1H), 6.73 (dd, J=8.0, 0.8 Hz, 1H), 6.83 (dd, J=7.6, 1.2 Hz, 1H),7.05 (ddd, J=7.6, 7.6, 1.2 Hz, 1H), 7.22 (ddd, J=7.6, 7.6, 1.2 Hz, 1H),7.30-7.39 (m, 2H), 7.90 (dd, J=7.6, 1.6 Hz, 1H). ³C NMR (400 MHz, CDCl₃)8.33.6, 46.4, 52.5, 116.7, 118.6, 123.2, 127.0, 128.3, 128.6, 128.8,129.9, 130.5, 134.6, 136.3, 144.6, 170.9. ESMS m/z 142.2, 322.4 [M+H]⁺.LC/MS t_(R) 8.85 (216.1 [M−HSCH₂CO₂CH₃+H]⁺, 322.1 [M+H]⁺) min.

11s (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11s was preparedand isolated as a white solid (1.84 g, 5.18 mmol, 70% yield). R_(f)(silica, 40-60 petroleum ether:ethyl acetate (2:1))=0.30. HPLC (214 nm)t_(R)=9.07 (91.4%) min. ¹H NMR (400 MHz, CDCl₃) δ 3.13 (d, J=16 Hz, 1H),3.21 (d, J=16 Hz, 1H), 3.73 (s, 3H), 4.39 (brs, 2H), 5.79 (s, 1H), 6.66(d, J=8.4 Hz, 1H), 6.81 (d, J=2.8 Hz, 1H), 7.04 (dd, J=8.4, 2.8 Hz, 1H),7.26-7.32 (m, 1H), 7.37-7.43 (m, 2H), 7.89-7.93 (m, 1H). ¹³C NMR (400MHz, CDCl₃) δ 33.6, 46.1, 52.6, 118.1, 123.8, 125.2, 127.2, 128.1,128.5, 129.2, 130.1, 130.3, 134.7, 135.4, 142.6, 170.9. ESMS m/z 250.2[M−HSCH₂CO₂CH₃+H]⁺, 356.3 [M+H]⁺. LC/MS t_(R) 9.41 (250.2[M−HSCH₂CO₂CH₃+H]⁺, 356.0 [M+H]⁺).

11t (W₁=5-nitro, R₁=phenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11b, 11t (486 mg, 1.46mmol, 67% overall yield for two steps) was obtained as a yellow oil:R_(f) (petroleum ether:ethyl acetate (2:1)=0.29). HPLC (214 nm)t_(R)=8.37 (96.4%) min. ¹H NMR (400 MHz, CDCl₃) δ 3.15 (d, J=17.2 Hz,1H), 3.23 (d, J=17.2 Hz, 1H), 3.74 (s, 3H), 5.35 (s, 1H), 5.38 (br s,1H), 6.68 (d, J=8.8 Hz, 1H), 7.33-7.46 (m, 3H), 7.51-7.55 (m, 2H), 7.81(d, J=2.4 Hz, 7.98 (dd, J=8.8, 2.4 Hz, 1H). ¹³C NMR (400 MHz, CDCl₃) δ33.1, 49.5, 52.6, 114.9, 122.9, 125.0, 125.9, 128.2, 129.0, 129.1,136.9, 138.6, 151.2, 171.5. ESMS m/z 227.2 [(M−HSCH₂CO₂CH₃+H)]⁺, 333.3[(M+H)]⁺. LC/MS t_(R) 9.25 (227.2 [(M−HSCH₂CO₂CH₃+H)]⁺, 333.1 [(M+H)]⁺,665.4 [2M+H]⁺) min.

11u (W₁=5-Cl, R₁=2-thiazolyl, R₂=NH₂)

Using a similar procedure as 11b, compound 11u was prepared and isolatedas a brown gum (62 mg, 18.4%), clogP=2.93; R_(f) (petroleum ether:ethylacetate (1:1))=0.35; HPLC (214 nm) t_(R)=7.58 (81%) min; ¹H NMR (400 mHz, CDCl₃) δ 3.27 (d, J=16.0 Hz, 1H), 3.35 (d, J=16.0 Hz, 1H), 3.72 (s,3H), 5.73 (s, 1H), 6.63 (d, J=8.8 Hz, 1H), 7.04-7.06 (m, 2H), 7.34 (d,J=3.3 Hz, 1H), 7.77 (d, J=3.3 Hz, 1H). ¹³C NMR (100 m Hz, CDCl₃) δ 33.4,47.3, 52.6, 118.0, 120.4, 123.0, 123.6, 128.5, 129.1, 143.0, 143.8,169.8, 170.6; ESMS mnz 329.4 [M+H]⁺.

11v (W₁=H, R₁=2,4-dimethylphenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, 11v was preparedand isolated as an off white solid (56 mg, 66% yield); clogP=3.96; R_(f)(petroleum ether:ethyl acetate (4:1)=0.70; HPLC (214 nm) t_(R)=9.17(95%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.14 (s, 3H), 2.32 (s, 3H), 3.13(d, J=15.9 Hz, 1H), 3.18 (d, J=15.9 Hz, 1H), 3.70 (s, 3H), 4.05 (br s,2H), 5.58 (s, 1H), 6.62-6.66 (m, 1H), 6.73 (dd, J=1.1, 7.9 Hz, 1H), 6.86(dd, J=1.4, 7.9 Hz, 1H), 6.98 (s, 1H), 7.04 (dd, J=1.4, 7.5 Hz, 1H),7.08 (d, J=7.5 Hz, 1H), 7.69 (d, J=7.9 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃)δ 18.9, 20.9, 33.5, 46.3, 52.4, 116.6, 118.8, 124.2, 126.8, 128.3,128.5, 128.8, 131.6, 133.3, 137.1, 144.7, 171.5; ESMS m/z 210.2[M−C₃H₄O₂S]⁺, 316.4 [M+H]⁺; LC/MS t_(R)=9.54 (210.1 [M−C₃H₅O₂S]⁺, 316.1[M+H]⁺, desired product 95%) min.

11w (W₁=5-Cl, R₁=2,4-dimethylphenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, 11w was preparedand isolated as an off white solid (89 mg, 84% yield); clogP=4.51; R_(f)(petroleum ether:ethyl acetate (4:1)=0.69; HPLC (214 nm) t_(R)=9.90(97%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.13 (s, 3H), 2.33 (s, 3H), 3.12(d, J=16.1 Hz, 1H), 3.18 (d, J=16.1 Hz, 1H), 3.71 (s, 3H), 4.29 (br s,2H), 5.52 (s, 1H), 6.63 (d, J=8.4 Hz, 1H), 6.83 (d, J=2.5 Hz, 1H),6.99-7.02 (m, 2H), 7.09 (d, J=7.5 Hz, 1H), 7.65 (d, J=7.9 Hz, 1H); ³CNMR (100 MHz, CDCl₃) δ 18.9, 20.9, 33.4, 46.0, 52.5, 117.5, 123.2,125.7, 127.1, 128.2, 128.3, 128.5, 131.8, 132.6, 137.0, 137.4, 143.7,171.4; ESMS m/z 244.3 [M−C₃H₄O₂S]⁺, 350.3 [M+H]⁺; LC/MS t_(R) _(=11.01)(244.0 [M−C₃H₄O₂S]⁺, 350.0 [M+H]⁺, desired product 95%) min.

11x (W₁=5-Cl, R₁=phenyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11c, 11x (1.18 g, 3.67mmol, 86% overall yield) as a yellow oil (R_(f) (40-60 petroleumether:ethyl acetate (2:1))=0.30). HPLC (214 nm) t_(R)=8.95 (94.4%) min.¹H NMR (400 MHz, CDCl₃) δ 3.13 (d, J=1.6 Hz, 1H), 3.20 (d, J=1.6 Hz,1H), 3.73 (s, 3H), 5.40 (s, 1H), 6.64 (d, J=8.4 Hz, 1H), 6.93 (d, J=2.8Hz, 1H), 7.04 (dd, J=8.4, 2.8 Hz, 1H), 7.30-7.35 (m, 1H), 7.37-7.43 (m,3H), 7.50 (d, J=7.6 Hz, 2H). ¹³C NMR (400 MHz, CDCl₃) δ 33.3, 49.7,52.5, 117.6, 123.0, 125.7, 127.8, 128.3, 128.8, 128.9, 129.0, 137.8,143.5, 171.1. ESMS m/z 216.2 [(M−HSCH₂CO₂CH₃+H)]⁺, 322.1 [(M+H)]⁺. LC/MSt_(R) 9.10 (216.3 [(M−HSCH₂CO₂CH₃+H)]⁺, 322.1 [(M+H)]⁺, 643.1 [(2M+H)]⁺)min.

11y (W₁=2-CH₃, R₁=2-amino-3-pyridinyl, R₂=H)

Using a similar procedure as for the synthesis of 11b, 11v was preparedand isolated as pale yellow crystals. HPLC (214 nm) t_(R)=6.08 min(92.1%). ¹H NMR (CDCl₃, 400 MHz) δ 2.15 (s, 3H), 3.17 (dd, J=26.0, 16.0Hz), 3.73 (s, 3H), 5.27 (bs, NH₂, 2H), 5.49 (s, 1H), 6.54 (dd, J=7.6,4.8 Hz, 1H), 7.04 (dd, J=7.6, 1.6 Hz, 1H), 7.17-7.32 (m, 3H), 7.83 (d,J=7.2 Hz, 1H), 7.98 (dd, J=4.8, 1.6 Hz, 1H). LCMS t_(R)=6.18 min (303,[M+H]⁺).

11aa (W₁=5-Cl, R₁=2-methylphenyl, R₂=NH₂)

From 10aa, isolated as a yellow powder (254 mg, 0.756 mol, 74% yield).R_(f) (petroleum ether:ethyl acetate (2:1)=0.60; MS=335.07(M+),(2M+1)⁺=671.4 (MS=335.07(M⁺), 671.4[2M+1]); ¹H NMR (400 MHz) 7.80 (1H,d, J=8.0 Hz), 7.07-7.32 (3H, m), 7.05 (1H, dd, J=2.4, 8.4 Hz), 6.93 (1H,d, J=8.4 Hz), ), 6.76 (1H, d, J=8.4 Hz), 5.56 (1H, s), 3.69 (3H, s),3.14-3.18 (2H, m) 2.15 (3H, s); ¹³CNMR (400 MHz) 171.42, 142.97, 137.29,135.53, 130.97, 128.49, 128.36, 127.80, 126.43, 125.99, 123.78, 117.97,52.56, 46.22, 33.40, 19.01.

11ab (W₁=5-Cl, R₁=2-methylphenyl, R₁=H)

From 10ab, isolated as a yellow oil (0.11 g, 71%). HPLC (214 nm)t_(R)=9.05 min (95.1%) ¹H NMR (400 MHz, CDCl₃) δ 2.30 (s, 3H), 2.36 (s,3H), 3.08 (s, 2H, NH₂), 3.65 (s, 3H), 5.58 (s, 1H), 7.02 (dd, J=6.4, 1.6Hz, 1H), 7.11-7.22 (m, 6H), 7.54 (d, J=7.2 Hz, 1H). LCMS t_(R)=9.87 min(318.1 [M+NH₄]⁺, 618.5 [2M+NH₄]⁺).

11ac (W₁=H, R₁=2-methylphenyl, R₂=methoxy) was obtained from 10ac as anoff-white solid (0.013 g, 32% yield). R_(f) (diethyl ether:petrol(1:4))=0.24. HPLC (214 nm) t_(R)=10.29 min (97.7%). ¹H NMR (CDCl₃, 400MHz) δ 2.36 (s, 3H), 3.12 (s, 2H), 3.62 (s, 3H), 3.78 (s, 3H), 5.99 (s,1H), 6.82 (d, J=8.4 Hz, 1H), 6.90-6.94 (m, 1H), 7.09-7.14 (m, 3H),7.17-7.21 (m, 1H) 7.47 (d, J=6.8 Hz, 1H), 7.52 (dd, J=7.6, 1.6 Hz, 1H).LCMS (214 nm) t_(R)=9.36 min (334.3 [M+NH₄]⁺, 376.3, 394.7, 527.3 (notidentified), 650.2 [2M+NH₄]⁺).

11ad (W₁=5-CH₃, R₁=2-methylphenyl, R₂=OH) was obtained from 10ad as acolorless oil (0.033 g, 100% yield). R_(f) (diethyl ether:petrol(1:2))=0.27. HPLC (214 nm) t_(R)=9.26 min (98.9%). ¹H NMR (CDCl₃, 400MHz) δ 2.21 (s, 3H), 3.20 (s, 2H), 3.76 (s, 3H), 5.72 (s, 1H), 6.75-6.79(m, 1H), 6.83 (dd, J=7.6, 1.6 Hz, 1H), 6.95 (dd, J=16, 0.8 Hz, 1H),7.13-7.29 (m, 5H), 7.79 (d, J=7.6 Hz, 1H). LCMS (214 nm) t_(R)=8.98 min(320.2 [M+NH₄]⁺, 499.0 (not identified), 622.2 [2M+NH₄]⁺).

11ae (W₁=H, R₁=5-amino-2-chlorophenyl, R₂=NH₂) was obtained from 10ae asyellow oil (158 mg, 72%). ¹H NMR (CDCl₃, 500 MHz) δ 3.14 (d, J=14.5 Hz,1H), 3.18 (d, J=14.5 Hz, 1H), 3.67 (s, 3H), 5.80 (s, 1H), 6.51 (dd,J=2.8, 8.7 Hz), 7.00 (d, J=2.8 Hz, 1H), 7.10 (d, J=9.7, 1H), 7.24 (m,1H), 7.31 (m, 2H), 7.42 (m, 2H).

11af (W₁=H, R₁=3-amino-4-chlorophenyl, R₂=NH₂) was obtained as yellowoil (142 mg, 88.5%). ¹H NMR (CDCl₃, 500 MHz) δ 3.09 (s, 2H), 3.68 (s,3H), 5.28 (s, 1H), 6.76 (dd, J=2.0, 8.2 Hz, 1H), 6.84 (d, J=2.0 Hz, 1H),7.17 (d, J=8.2 Hz, 1H), 7.25 (m, 1H), 7.31 (m, 2H), 7.40 (m, 2H).

11ag (W₁=5-Cl, R₁=2-chlorophenyl, R₂=—OH)

Compound 10ag (50 mg, 0.18 mmol) in dry CH₂Cl₂ (1 ml) was cooled to 10°C. To the cold stirred solution anhyd. ZnCl₂ (73 mg, 0.54 mmol) andmethyl thioglycolate (32 μl, 0.36 mmol) were added and stirring wascontinued overnight. The reaction mixture was quenched by adding water(4 ml), extracted with CH₂Cl₂, washed with water, dried over anhyd.sodium sulfate and concentrated to afford 60 mg of crude product. Theproduct was purified by column using 10% ethyl acetate in petroleumether to yield 48 mg (62%) of compound 11ag. m.p: 160.3-162.7° C. ¹H NMR(300 MHz, CDCl₃) δ 7.90 (d, J=8.4 Hz, 1H), 7.49 (s, phenolic-OH),7.39-7.26 (m, 2H), 7.13 (dd, J=2.1 Hz, 8.4 Hz, 2H), 6.92 (d, J=8.7 Hz,1H), 6.7 (d, J=2.1 Hz, 1H), 5.87 (s, 1H), 3.78 (s, 3H), 3.25 (d, J=16.8Hz, 1H), 3.18 (d, J=16.8, 1H). GC-MS calcd. for C₁₆H₁₄Cl₂O₃S: 356.00;found: 356/358 (M⁺), 251/253, 215/217, 181, 152.

11ah (W₁=5-Cl, R₁=phenyl, R₂=—OCH3)

Compound 10ah (25 mg, 0.116 mmol) in dry DCM (1 ml) was cooled to 0° C.To this cold stirred solution SnCl₄ (27 μl, 0.23 mmol) followed bymethyl thioglycolate (20 μL, 0.23 mmol) were added and stirring wascontinued overnight. The reaction was quenched by adding 1M HCl (2 ml)extracted with DCM (10 ml), washed with 1M HCl (2×10 ml) dried oversodium sulfate and concentrated. The crude product was purified by 10%EtOAc/PE to yield 24 mg (68%) of the product. ¹H NMR (300 MHz, CDCl₃) δ7.55 (d, J=1.5 Hz, 1H), 7.44 (d, J=7.5 Hz, 1H), 7.15-7.33 (m, 5H), 6.76(d, J=9 Hz, 1H), 5.77 (s, 1H), 3.79 (s, 3H), 3.65 (s, 3H), 3.13 (s, 2H).

11ai (W₁=5-Cl, R₁=phenyl, R₂=—OH)

Compound 10ai (200 mg, 0.83 mmol) in dry DCM (2 ml) was cooled to 0° C.To the cooled stirred solution anhydrous ZnCl₂ (681 mg, 5 mmol) andmethyl thioglycolate (223 μl, 2.5 mmol) were added and stirring wascontinued overnight. The reaction mixture was quenched by adding water(5 ml), extracted with DCM and washed with water. Dried over Na₂SO₄ andconcentrated to afford 224 mg (81%) of compound 11ai. m.p: 123-125.3° C.¹H NMR (300 MHz, CDCl₃) δ 7.39 (d, J=7.2 Hz, 1H), 7.32-7.20 (m, 4H),7.038 (dd, J=8.4 Hz, 2.6 Hz, 1H), 6.84 (d, J=2.1 Hz, 1H), 6.79 (d, J=8.4Hz, 1H), 5.45 (s, 1H), 3.67 (s, 3H), 3.12 (s, 2H).

Synthesis of 11aj (W₁=5-Cl, R₁=2-chlorophenyl, R₂=H)

To a mixture of alcohol 10aj (500 mg, 1.98 mmol) in 1 mL of TFA wasadded methyl thioglycolate (550 mg, 5.18 mmol). The reaction mixture wasstirred at room temperature for 3 days TFA was removed under highvacuum, the residue diluted with DCM, washed with NaHCO₃, dried overanhyd. Na₂SO₄ and concentrated to afford 11aj (600 mg). ¹H NMR (300 MHz,CDCl₃) δ 7.65 (dd, J=7.5 Hz, J=1.5 Hz, 1H), 7.42 (s, 1H), 7.21-7.38 (m,6H), 5.86 (s, 1H), 3.67 (s, 3H), 3.17 (d, J=15 Hz, 1H), 3.12 (d, J=15Hz, 1H).

Synthesis of 11ak (W₁=5-Cl, R₁=cyclohexyl, R₂=NH₂)

Alcohol 10ak (0.25 g, 0.735 mmol) was mixed with methyl thioglycolate(0.656 mL, 7.35 mmol, 10 equivalents) and TFA (1.13 mL, 14.7 mmol, 20equivalents). The mixture was stirred for 12 hrs. The TFA was thenevaporated under vacuum and the resulting solution diluted with CH₂Cl₂(20 mL), washed with a saturated solution of NaHCO₃ (2x10 mL), water (10mL) and brine (20 mL). The organic layer was dried over sodium sulfate,filtered and concentrated under vacuum. The crude product was purifiedby flash chromatography on a silica gel column using a mixture ofpetroleum ether and ethyl acetate as eluent to give 11ak as colorlessoil (0.19 g, 80% yield); LC/MS calcd. for C₁₆H₂₂ClNO₂S 327 [M−C₃H₅O₂S]⁺,found: 222.

Synthesis of 11al (W₁=5-Cl, R₁=iso-propyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11ak, 11al wasprepared from 10al as a colorless oil (88% yield); LC/MS calcd. forC₁₃H₁₈ClNO₂S: 287 [M−C₃H₅O₂S]⁺, found: 182.

Synthesis of 11am (W₁=5-Cl, R₁=tert-butyl, R₂=NH₂)

Using a similar procedure as for the synthesis of 11ak, 11 am wasprepared from 10am as a colorless oil (86% yield); LC/MS calcd. forC₁₄H₂₀ClNO₂S: 301 [M−C₃H₅O₂S]⁺, found: 196.

Synthesis of 11an (W₁=5-Cl, R₁=—(CH₃)₃OCH₂(C₆H₅), R₂=NH₂)

Using a similar procedure as for the synthesis of 11ak, llan wasprepared from 10 am as a colorless oil (78% yield); LC/MS calcd. forC₂₀H₂₄ClN₀₃S: 393 [M−C₃H₅O₂S]⁺, found: 288.

Synthesis of Thioether Carboxylic Acid 12 12t (W₁=5-nitro, R₁=phenyl,R₂=NH₂)

To a stirred solution of methyl ester 11t (39.3 mg, 0.118 mmol) in THF(2.4 ml) and methanol (2.4 ml) at rt was added sodium hydroxide solution(1.0 M x 2.4 ml, 20eq). After stirring for 30 min. the reaction mixturewas partitioned between brine and dichloromethane. The aqueous phase wastitrated to exactly pH 7.0 with concentrated hydrochloric acid, andextracted twice with dichloromethane. The combined organic phase wasdried with brine and sodium sulfate, then filtered and evaporated togive carboxylic acid 12t (38.0 mg, 0.119 mmol, 100% yield) as a yellowgum, which is analytically pure. R_(f) (silica, dichloromethane:methanol (9:1))=0.25. HPLC (214 nm) t_(R)=7.62 (91.4%) min. ¹H NMR (400MHz, CDCl₃) δ 3.19 (d, J=16.8 Hz, 1H), 3.27 (d, J=16.8 Hz, 1H), 5.37 (s,1H), 6.60-7.10 (br s, 3H), 6.68 (d, J=8.8 Hz, 1H), 7.36 (d, J=7.2 Hz,1H), 7.43 (dd, J=7.2, 7.2 Hz, 1H), 7.52 (d, J=7.2 Hz, 1H), 7.91 (d,J=2.4 Hz, 1H), 8.00 (dd, J=8.8, 2.4 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) δ33.1, 49.7, 115.2, 122.6, 125.2, 126.1, 128.4, 128.9, 129.1, 136.6,138.7, 151.0, 176.2. ESMS m/z 227.2 [(M−HSCH₂CO₂H+ H)]⁺, 319.4 [(M+H)]⁺.LC/MS t_(R) 7.94 (227.0 [(M−HSCH₂CO₂H+ H)]⁺, 318.9 [(M+H)]⁺, 637.1[(2M+H)]⁺, 955.3 [3M+H]⁺) min.

Using a similar procedure, the following thioether carboxylic acids wereprepared from corresponding methyl esters.

12s (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂): white solid (657 mg, 1.92mmol, 100% yield). R_(f) (silica, dichloromethane:methanol (9:1))=0.25.HPLC (214 nm) t_(R)=8.27 (94.2%) min. ¹H NMR (400 MHz, CDCl₃) δ 3.20 (d,J=16 Hz, 1H), 3.22 (d, J=16 Hz, 1H), 6.02 (s, 1H), 7.07 (d, J=2.4 Hz,1H), 7.31-7.37 (m, 2H), 7.40-7.47 (m, 2H), 7.80-7.89 (m, 2H), 9.02 (s,1H). ³C NMR (400 MHz, CDCl₃) δ 32.8, 45.0, 126.1, 127.6, 129.0, 129.8,129.9, 130.5, 131.7, 132.6, 132.7, 133.9, 134.6, 175.0. ESMS m/z 250.2[M−HSCH₂CO₂H+ H]⁺, 342.1 [M+H]⁺. LC/MS t_(R) 8.66 (346.0[M−HSCH₂CO₂H+CF₃CO+H]⁺, 438.1 [M+CF₃CO+H]⁺, 874.8 [2(M+CF₃CO)+H]⁺.

12x (W₁=5-Cl, R₁=phenyl, R₂=NH₂) (46.0 mg, 0.149 mmol, 77% yield) as atan solid. R_(f) (silica, dichloromethane:methanol (9:1))=0.25. HPLC(214 nm) t_(R)=7.61 (90.3%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.79-3.14 (m,2H), 5.27 (s, 1H), 5.90-6.30 (br s, 3H), 6.48 (d, J=8.0 Hz, 1H), 6.85(s, 1H), 6.95 (d, J=8.0 Hz, 1H), 7.25-7.35 (m, 2H), 7.35-7.45 (m, 2H).¹³C NMR (100 MHz, CDCl₃) δ 34.8, 49.4, 118.7, 124.6, 127.4, 128.0,128.3, 128.7, 128.8, 129.0, 137.4, 141.8, 176.4. ESMS nvz 308.2 [M+H]⁺,349.2 [M+CH₃CN+H]⁺, 615.1 [2M+H]. LC/MS t_(R) 7.89 (216.0[M−HSCH₂CO₂H+H]⁺, 308.2 [M+H]⁺, 615.1 [2M+H]⁺, 921.9 [3M+H]⁺) min.

12r (W₁=H, R₁=2-chlorophenyl, R₂=NH₂)

From 11r, 12r was obtained as a white powder. HPLC (214 nm) t_(R)=7.33(85.8%) min. ¹H NMR (400 MHz, CDCl₃) δ 3.16 (d, J=16.0 Hz, 1H), 3.23 (d,J=16.0 Hz), 5.87 (s, 1H), 6.14 (br s, 1H), 6.71 (dd, J=3.2, 3.2 Hz, 1H),6.78 (d, J=7.6 Hz, 1H), 6.84 (d, J=7.6 Hz, 1H), 7.10 (ddd, J=8.0, 8.0,1.2 Hz, 1H), 7.25-7.31 (m, 1H), 7.34-7.42 (m, 1H), 7.97 (d, J=7.6 Hz,1H). ESMS m/z 308.5 [(M+H)]⁺. LC/MS t_(R) 7.06 (308.1 [(M+H)]⁺) min.

Synthesis of Thioether Carboxamides 13 13a (W₁=5-Cl, R₁=2-chlorophenyl,R₂=NH₂, R_(3a)=CH₃, R_(3b)=H)

To carboxylic acid 12s (115 mg, 0.337 mmol) under a nitrogen atmosphereat rt was added methylamine (2 mol/L in THF, 1.68 ml, 3.37 mmol, 10eq),EDC (129 mg, 0.673 mmol, 2eq) and DMAP (4.1 mg, 0.033 mmol, 0.1 eq) andthe resulting solution was stirred for 18 hrs. The reaction mixture wasevaporated and the residue partitioned between brine anddichloromethane. The aqueous phase was extracted with furtherdichloromethane and the combined organic extracts were dried with brineand sodium sulfate, then filtered and evaporated to give the crude amide(149 mg) as a yellow oil. The crude material was purified by flashchromatography on silica (5 g) with 40-60 petroleum ether:ethyl acetate(1:1 then 2:1) to give amide 13a (74.0 mg, 0.208 mmol, 62%) as a whitesolid. R_(f) (silica, petroleum ether:ethyl acetate (1:1))=0.15. HPLC(214 nm) t_(R)=8.54 (96.4%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.81 (d,J=4.8 Hz, 3H), 3.04-3.16 (m, 2H), 4.17 (br s, 2H), 5.64 (s, 1H), 6.40(br s, 1H), 6.62 (d, J=8.0 Hz, 1H), 7.01-7.07 (m, 2H), 7.26 (ddd, J=5.6,1.6, 1.6 Hz, 1H), 7.33 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 7.39 (dd, J=7.6,1.6, Hz, 1H), 7.72 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) δ26.5, 35.6, 46.1, 117.7, 123.1, 124.6, 127.4, 128.0, 128.4, 129.1,129.9, 130.2, 134.0, 136.0, 143.5, 168.9. ESMS m/z 355.1 [M+H]⁺. LC/MSt_(R)=7.80 (250.1 [M−HSCH₂CONHMe+H]⁺, 355.0 [M+H]⁺, 709.0 [2M+H]⁺) min.

13b (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=CH₃, R_(3b)=CH₃)

Using a similar procedure as for the synthesis of 13a, 13b was preparedfrom 12s and dimethylamine, and obtained as a white solid (123 mg, 0.333mmol, 99%). R_(f) (silica, petroleum ether:ethyl acetate (1:1))=0.20.HPLC (214 nm) t_(R)=9.10 (100%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.91 (s,3H), 2.95 (s, 3H), 3.15-3.27 (m, 2H), 4.69 (br s, 2H), 5.76 (s, 1H),6.62 (d, J=8.4 Hz, 1H), 6.75 (d, J=2.4 Hz, 1H), 6.99 (dd, J=8.4, 2.4 Hz,1H), 7.21-7.29 (m, 1H), 7.34-7.40 (m, 2H), 7.93-7.98 (m, 1H). ¹³C NMR(100 MHz, CDCl₃) δ 33.3, 36.0, 37.4, 46.0, 117.2, 122.2, 124.7, 127.0,127.9, 128.2, 128.9, 129.9, 130.4, 134.7, 136.2, 144.2, 168.7. ESMS m/z369.2 [(M+H)]⁺. LC/MS t_(R)=8.28 (368.9 [(M+H)]⁺, 737.1 [(2M+H)]⁺) min.

13c (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=2-phenylethyl, R_(3b)=H)

To a solution of 11s (52 mg, 0.15 mmole) in methanol (0.5 ml) was addedphenethylamine (0.19 ml, 1.5 mmole). The mixture was heated at 70° C.for 16 hrs. TLC indicated complete consumption of 11s. The solvent wasremoved under vacuum. The residue was dissolved in DCM (10 ml), washedwith 10% citric acid (3×10 ml) and dried over anhydrous sodium sulfate.The crude product was purified on a silica gel column using 20% ethylacetate in hexane as eluent to give 13c as white solid (60.4 mg, 90%yield). ¹H NMR (500 MHz, CDCl₃) δ 2.83 (t, J=7.0 Hz, 2H), 3.05 (d, J=16Hz, 1H), 3.10 (d, J=16 Hz, 11H), 3.61-3.48 (m, 2H), 4.10 (br, s, 2H),5.60 (s, 11H), 6.29 (br, s, 11H), 6.61 (d, J=8.4 Hz, 1H), 6.99 (d, J=2.2Hz, 1H), 7.03 (dd, J=8.3, 2.0 Hz, 1H), 7.18 (d, J=7.0 Hz, 1H), 7.32-7.21(m, 6H), 7.38 (d, J=7.0 Hz, 1H), 7.72 (d, J=7.5 Hz, 1H). LC-MS: calcd.For C₂₃H₂₂Cl₂N₂OS: 444.1; found: 444.8 [M+H]⁺.

Using a similar procedure as for the synthesis of 13c, the followingthioether amide was prepared.

13d (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a), R_(3b)=—(CH₂)₅—)

¹H NMR (500 MHz, CDCl₃) δ 1.65-1.50 (m, 6H), 3.19 (d, J=15 Hz, 1H), 3.25(d, J=15 Hz, 1H), 3.30-3.27 (m, 2H), 3.60-3.50 (m, 2H), 4.69 (br, s,2H), 5.74 (s, 1H), 6.63 (d, J=9.0 Hz, 1H), 6.76 (d, J=2.7 Hz, 1H), 7.00(dd, J=8.3, 2.1 Hz, 1H), 7.28 (dd, J=8.9, 1.8 Hz, 1H), 7.38 (t, J=7.6Hz, 2H), 7.95 (d, J=7.5 Hz, 1H). LC-MS: calcd. For C₂₀H₂₂Cl₂N₂OS: 408.1;found: 408.9 [M+H]⁺.

13e (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=2-hydroxyethyl,R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 3.12 (d, J=16 Hz, 1H), 3.17 (d, J=16 Hz, 1H),3.44-3.36 (m, 1H), 3.55-3.48 (m, 1H), 3.82-3.72 (m, 2H), 4.10 (br, s,2H), 5.66 (s, 1H), 6.65 (d, J=9.0 Hz, 1H), 6.80 (br, s, 1H), 7.06-7.03(m, 2H), 7.28 (d, J=7.1 Hz, 1H), 7.35 (t, J=8.1 Hz, 1H), 7.40 (d, J=8.3Hz, 1H), 7.75 (d, J=8.2 Hz, 1H). LC-MS: calcd. For C₁₇H₁₈Cl₂N₂O₂S:384.0; found: 384.8 [M+H]⁺.

13f (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=benzyl, R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 3.14 (d, J=16 Hz, 1H), 3.18 (d, J=16 Hz, 1H),4.45-4.38 (m, 1H), 4.55-4.48 (m, 1H), 4.10 (br, s, 2H), 5.60 (s, 1H),6.60 (d, J=8.7 Hz, 1H), 6.72 (br, s, 1H), 7.03 (d, J=9.1 Hz, 2H),7.37-7.23 (m, 8H), 7.71 (d, J=9.0 Hz, 1H). LC-MS: calcd. ForC₂₂H₂₀Cl₂N₂OS: 430.1; found: 430.8 [M+H]⁺.

13g (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=(2-acetamido)ethyl,R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 1.95 (s, 3H), 3.07 (d, J=16 Hz, 1H), 3.14 (d,J=16 Hz, 1H), 3.44-3.35 (m, 4H), 4.10 (br, s, 2H), 6.18 (br, s, 1H),6.64 (d, J=8.3 Hz, 1H), 6.93 (br, s, 1H), 6.97 (d, J=2.0 Hz, 1H), 7.04(dd, J=8.3, 2.0 Hz, 1H), 7.28 (d, J=7.3 Hz, 1H), 7.36 (t, J=7.2 Hz, 1H),7.39 (d, J=8.3 Hz, 1H), 7.78 (d, J=8.1 Hz, 1H). LC-MS: calcd. ForC₁₉H₂₁Cl₂N₃O₂S: 425.07; found: 425.9 [M+H]⁺.

13h (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=cyclohexyl, R_(3b)=H)

¹H NMR (500 MHz, CDCl₃)δ 1.98-1.12 (m, 10H), 3.06 (d, J=16 Hz, 1H), 3.11(d, J=16 Hz, 1H), 3.84-3.75 (m, 1H), 4.10 (br, s, 2H), 5.63 (s, 1H),6.16 (br, d, J=7.9 Hz, 1H), 6.64 (d, J=9.1 Hz, 1H), 7.05-7.02 (br, m,2H), 7.27 (d, J=7.3 Hz, 1H), 7.34 (t, J=7.3 Hz, 1H), 7.40 (d, J=7.9 Hz,1H), 7.73 (d, J=7.9 Hz, 1H). LC-MS: calcd. For C₂₁H₂₄Cl₂N₂OS: 422.1;found: 422.9 [M+H]⁺.

13i (W₁=5-Cl, R₁=2-chlorophenyl, R₂NH₂R_(3a), =2,2-diphenylethyl,R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 3.00 (d, J=16 Hz, 1H), 3.04 (d, J=16 Hz, 1H),3.90-3.85 (m, 1H), 3.98-3.94 (m, 1H), 4.08 (br, s, 1H), 4.10 (br, s,2H), 5.55 (s, 1H), 6.21 (br, s, 1H), 6.60 (d, J=8.4 Hz, 1H), 6.91 (d,J=2.1 Hz, 1H), 7.02 (dd, J=8.4, 2.2 Hz, 1H), 7.34-7.20 (m, 12H), 7.38(d, J=8.1 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H). LC-MS: calcd. ForC₂₉H₂₆Cl₂N₂OS: 520.1; found: 520.9 [M+H]⁺.

13j (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH—, R_(3a),R_(3b)=—(CH₂)₂—N(Ph)-(CH₂)₂—)

¹H NMR (500 MHz, CDCl₃) δ 3.18-3.10 (m, 4H), 3.25 (d, J=16 Hz, 1H), 3.30(d, J=16 Hz, 1H), 3.56-3.52 (br, m, 2H), 3.82-3.78 (br, m, 2H), 4.56(br, s, 2H), 5.76 (s, 1H), 6.64 (d, J=8.6 Hz, 1H), 6.85 (d, J=2.0 Hz,1H), 6.95-6.90 (m, 3H), 7.02 (dd, J=8.0, 2.0 Hz, 1H), 7.32-7.25 (m, 3H),7.37 (t, J=7.7 Hz, 2H), 7.90 (d, J=7.7 Hz, 1H). LC-MS: calcd. ForC₂₅H₂₅Cl₂N₃OS: 485.1; found: 485.9 [M+H]⁺.

13k (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=n-propyl, R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 0.93 (t, J=7.4 Hz, 3H), 1.58-1.50 (m, 2H),3.10 (d, J=16 Hz, 1H), 3.15 (d, J=16 Hz, 1H), 3.28-3.20 (m, 2H), 4.20(br, s, 2H), 5.63 (s, 1H), 6.33 (br, s, 1H), 6.63 (d, J=8.5 Hz, 1H),7.06-7.02 (m, 2H), 7.30-7.25 (m, 1H), 7.33 (t, J=7.2 Hz, 1H), 7.40 (d,J=7.5 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H). LC-MS: calcd. For C₁₈H₂₀Cl₂N₂OS:382.1; found: 382.8 [M+H]⁺.

13l (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=n-hexyl, R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ 0.88 (t, J=5.8 Hz, 3H), 1.36-1.26 (br, m, 6H),1.55-1.46 (m, 2H), 3.10 (d, J=16 Hz, 1H), 3.14 (d, J=16 Hz, 1H),3.29-3.22 (m, 2H), 4.20 (br, s, 2H), 5.63 (s, 1H), 6.30 (br, s, 1H),6.64 (d, J=8.5 Hz, 1H), 7.06-7.02 (m, 2H), 7.30-7.25 (m, 1H), 7.33 (t,J=7.2 Hz, 1H), 7.40 (d, J=7.5 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H). LC-MS:calcd. For C₂₁H₂₆Cl₂N₂OS: 424.1; found: 424.9 [M+H]⁺.

13m (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=3-ethoxypropyl,R_(3b)=H)

LC-MS: calcd. For C₂₀H₂₄Cl₂N₂O₂S: 426.1; found: 426.9 [M+H]⁺.

13n (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=cyclohexylmethyl,R_(3b)=H

LC-MS: calcd. For C₂₂H₂₆Cl₂N₂OS: 436.1; found: 437.0 [M+H]⁺.

13o (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R=propargyl, R_(3b)=H)

LC-MS: calcd. For C₁₈H₁₆Cl₂N₂OS: 378.0; found: 378.8 [M+H]⁺.

13p (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=3-(1-imidazolyl)propyl,R_(3b)=H)

¹H NMR (500 MHz, CDCl₃) δ2.05-1.98 (m, 2H), 3.10 (d, J=16 Hz, 1H), 3.15(d, J=16 Hz, 1H), 3.34-3.24 (m, 2H), 3.99 (t, J=6.8 Hz, 2H), 4.20 (br,s, 2H), 5.61 (s, 1H), 6.42 (br, s, 1H), 6.63 (d, J=8.5 Hz, 1H), 6.93 (s,1H), 7.08-7.03 (m, 2H), 7.11 (d, J=2.7 Hz, 1H), 7.28 (d, J=7.5 Hz, 1H),7.34 (t, J=6.9 Hz, 1H), 7.40 (d, J=8.1 Hz, 1H), 7.49 (s, 1H), 7.66 (d,J=8.9 Hz, 1H). LC-MS: calcd. For C₂₁H₂₂Cl₂N₄OS: 448.1; found: 449.0[M+H]⁺.

13q (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R_(3a)=3-(N,N-dimethylamino)propyl, R_(3b)=H)

LC-MS: calcd. For C₂₀H₂₅Cl₂N₃OS: 425.1; found: 426.1 [M+H]⁺.

13r (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂=cyclopropyl, R_(3b)=H)

LC-MS: calcd. For C₁₈H₁₈Cl₂N₃OS: 380.0; found: 380.9 [M+H]⁺.

13s (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R_(3a)=2-(4-hydroxyphenyl)ethyl, R_(3b)=H)

LC-MS: calcd. For C₂₃H₂₂Cl₂N₂O₂S: 460.08; found: 460.9.

Synthesis of 13t (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R_(3a)=2-(3-iodo-4-hydroxyphenyl)ethyl, R_(3b)=H) and 13u (W₁=5-Cl,R₁=2-chlorophenyl, R₂=NH₂, R_(3a)=2-(3,5-diiodo-4-hydroxyphenyl)ethyl,R_(3b)=H)

13s (23 mg, 50 μmole) was dissolved in ethanol (50 ml). De-ionized water(200 ml) was added and the solution became slightly cloudy. NaI (20 mg,200 tmole), 30% hydrogen peroxide (1.0 ml) and lactoperoxidase (LPO)(100 μl×1 unit/μl) were added. The mixture was shaken gentlycontinuously at room temperature on a shaker. The progress of thereaction was monitored using LC-MS. After 3 hrs, the reaction mixturebecame very cloudy. LC-MS indicated about 15% conversion. The reactionmixture was diluted with 20% aqueous ethanol (250 ml) upon which thereaction mixture became clear. LPO (250 μl×1 unit/μl) was added and themixture was shaken over night. LC-MS showed about 50% conversion and theformation of small amount of di-iodo-product. The ethanol was removedunder vacuum. The aqueous residue was extracted with dichloromethane (50ml×4). The combined organic layer was dried over anhydrous sodiumsulfate. The crude product was dissolved in methanol (3 ml) and purifiedon reverse phase preparative HPLC. 13t: white solid (10.6 mg). LC-MS:calcd. For C₂₃H₂₁Cl₂₁N₂O₂S: 585.97; found: 586.8. 13u: white solid (3.1mg). LC-MS: calcd. For C₂₃H₂OCl₂I₂N₂O₂S: 711.87; found: 712.7.

13v (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R_(3a)=—(CH₂)₂O(CH₂)₂O(CH₂)₂NH₂, R_(3b)=H)

Yellow viscous oil (0.505 g, quantitative). LC-MS: calcd. ForC₂₃H₂₇Cl₂N₃O₃S: 471.12; found: 472.1.

13ag (W₁=5-Cl, R₁=2-chlorophenyl, R₂=OH, R_(3a) & R_(3b)=—(CH₂)₅—)

m.p.=219.2-221.2° C. ¹H NMR (300 MHz, CDCl₃) δ 9.7 (s, phenolic-OH),8.01 (d, J=7.2 Hz, 1H), 7.37-7.44 (m, 2H), 7.31 (d, J=6.6 Hz, 1H) 7.09(dd, J=2.7 Hz, 9 Hz, 1H), 6.94 (dd, J=8.7 Hz, 1H), 6.6 (d, J=2.4 Hz,1H), 5.84 (s, 1H), 3.64 (s, 2H), 3.35-3.36 (m, 2H), 3.26 (d, J=15.9 Hz,1H), 3.18 (d, J=15.9 Hz, 1H), 1.55-1.59 (m, 6H).

GC-MS: calcd. for C₂₀H₂₁Cl₂NO₂S: 409.07; found: 410 (MH⁺), 215/217,152,126/127, 112.

13ai (W₁=5-Cl, R₁=phenyl, R₂=OH, R_(3a) & R_(3b)=—(CH₂)₅—

m.p.=192° C. ¹H NMR (300 MHz, CDCl₃) δ 9.58 (s, 1H), 7.48 (d, J=7.8 Hz,1H) 7.415-7.30 (m, 3H), 7.1-7.06 (dd, J=8.4 Hz, 3 Hz, 2H), 6.92 (d, J=9Hz, 1H), 6.76 (d, J=2.1 Hz, 1H), 5.48 (s, 1H), 3.72-3.53 (m, 4H), 3.16(d, J=16.2 Hz, 1H), 3.27 (d, J=16.2 Hz, 1H), 1.64-1.61 (m, 6H).

13aj (W₁=5-Cl, R₁=2-chlorophenyl, R₂=H, R_(3a)=cyclohexylmethyl,R_(3b)=H)

Compound 13aj was obtained from 11aj and cyclohexylmethylamine as awhite solid. LC-MS: calcd. for C₂₂H₂₅Cl₂NOS: 421.10, found: 421.9(M+H)⁺.

EXAMPLE 6

14a (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, O=4-fluorophenyl)

To the solution of 12s (0.342 g, 1 mmol) in diglyme (3 ml) was added4-fluorobenzamidoxime (from Aldrich, 0.154 g, 1 mmol) and EDC (0.38 g, 2mmol). The mixture was stirred at 50° C. for 16 hrs followed by at 110°C. for 3 hrs. The crude product was purified on a silica gel columnusing 30% ethyl acetate in hexane to give 14a as a white solid (0.116 g,25%): ¹H NMR (400 MHz, CDCl₃) δ 3.78 (d, 1H), 3.86 (d, 1H), 6.00 (s,1H), 6.64 (d, 1H), 6.93 (d, 1H), 7.03 (dd, 1H), 7.19 (m, 2H), 7.27 (m,2H), 7.36 (m, 2H), 7.86 (d, 1H), 8.08 (m, 1H). LC-MS: calcd. forC₂₂H₁₆Cl₂FN₃OS: 459.04; found: 459.8.

14b (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, O=methyl)

Using a similar procedure as for 14a, 14b was obtained from 12s andacetamidoxime as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 2.39 (s, 3H),3.71 (d, J=15.8 Hz, 1H), 3.77 (d, J=15.8 Hz, 1H), 5.80 (s, 1H), 6.64 (d,J=8.4 Hz, 1H), 6.92 (d, J=2.4 Hz, 1H), 7.04 (dd, J=2.4, 8.4 Hz, 1H),7.28 (dd, J=7.8, 7.9 Hz, 1H), 7.38 (m, 2H), 7.81 (d, J=7.9 Hz, 1H).

EXAMPLE 7

General Procedures for the Synthesis of 14 from 10 and3-methyl-(5-mercantomethyl)-1,2,4-oxadiazole

Compound 10 was converted to 14 using one of the three alternativemethods listed below. The experimental detail for each compound wasnoted along with its spectral data.

(a) Compound 10 was dissolved in anhydrous dichloromethane (1 ml). The1-methyl-5-(mercaptomethyl)-1,2,4-oxadiazole (synthesized in five stepsfollowing a procedure from Broughton et al. published PCT WO98/04559) (1equivalent) was added followed by TFA (1 equivalent). The reactions weremonitored by TLC. After 1 hr the reaction mixture was diluted withdichloromethane (20 ml) and washed with 1:1 brine/1 M NaOH (10 ml). Theaqueous layer was extracted with dichloromethane (1×10 ml). The combinedorganic phase was dried over sodium sulfate, filtered and the solventremoved in vacuo. The crude material was purified by flashchromatography on silica gel (25 g) using petroleum spirit/ethyl acetateas eluent. The desired compound from the flash column was taken up into4 ml of 90% aqueous acetonitrile and was lyophilised to yield thedesired products.

(b) The procedure was essentially the same as (a) except a large excessof TFA (200 uL) was added to each reaction and the reactions were leftovernight.

(c) The procedure was essentially the same as (a) except neat TFA (1 ml)was used instead of the dichloromethane and the reaction was heated at60° C. over the weekend.

14d (W₁=5-Cl, R₁=3,5-dimethylphenyl, R₂=NH₂)

Compound 14d was synthesized using procedure (b) and obtained as agolden colored oil (55 mg, 66% yield); clogP=6.00; R_(f) (petroleumspirit/ethyl acetate, 2.5:1)=0.35; HPLC (214 nm) t_(R)=10.22 (96.79%)min; ¹H NMR (400 MHz, CDCl₃) δ 2.31 (s, 6H), 2.40 (s, 3H), 3.68 (d,J=15.9 Hz, 1H), 3.73 (d, J=15.9 Hz, 1H), 4.13 (brs, 2H), 5.35 (s, 1H),6.61 (d, J=8.2 Hz, 1H), 6.93 (s, 1H), 7.02-7.07 (m, 4H); t³C NMR (100MHz, CDCl₃) δ 11.6, 21.3, 26.1, 49.8, 117.8, 123.3, 125.5, 126.6, 128.5,129.0, 129.8, 137.3, 138.5, 143.4, 167.4, 176.9; LC/MS t_(R)=9.40 (374.3[M+H]⁺) min.

14g (W₁=5-Cl, R₁=2-thiophenyl R₂=NH₂)

Compound 14g was using procedure (a) and obtained as a golden coloredoil (31 mg, 34% yield); clogP=5.01; R_(f) (petroleum spirit/ethylacetate, 2:1)=0.3; HPLC (214 nm) t_(R)=9.27 (90.20%) min; ¹H NMR (400MHz, CDCl₃) δ 2.40 (s, 3H), 3.77 (d, J=16.1 Hz, 1H), 3.83 (d, J=16.1 Hz,1H), 4.22 (brs, 2H), 5.66 (s, 1H), 6.62 (d, J=8.5 Hz, 1H), 7.00 (dd,J=3.5, 5.1 Hz, 1H), 7.05 (dd, J=2.4, 8.5 Hz, 1H), 7.10 (brd, J=3.5 Hz,1H), 7.12 (d, J=2.4 Hz, 1H), 7.30 (dd, J=1.1, 5.1 Hz, 1H); ¹³C NMR (100MHz, CDCl₃) δ 11.5, 26.2, 45.1, 118.0, 123.3, 125.0, 126.1, 127.1,127.3, 128.8, 128.9, 141.9, 143.2, 167.4, 176.7; ESMS m/z 352.0 [M+H]⁺,393.2 [M+CH₃CN+H]⁺; LC/MS t_(R)=8.55 (351.9 [M+H]⁺) min.

14h (W₁=5-Cl, R₁=1-methyl-2-pyrrolyl, R₂=NH₂)

Compound 14h was synthesized using procedure (a) and obtained as a darkgreen solid (19 mg, 42% yield); clogP=3.81; R_(f) (petroleumspirit/ethyl acetate, 1:1)=0.70; HPLC (214 nm) t_(R)=9.06 (78.26%) min;¹H NMR (400 MHz, CDCl₃) δ 2.40 (s, 3H), 3.35 (s, 3H), 3.75 (d, J=16.1Hz, 1H), 3.90 (d, J=16.1 Hz, 1H), 4.05 (brs, 2H), 5.42 (s, 1H), 6.12 (t,J=3.2 Hz, 1H), 6.38-6.40 (m, 1H), 6.60-6.62 (m, 2H), 7.00 (d, J=2.4 Hz,1H), 7.03 (dd, J=2.4, 8.4 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 11.6,26.4, 33.7, 42.1, 107.2, 110.0, 117.9, 123.5, 123.8, 124.3, 128.0,128.7, 128.9, 143.3, 167.3, 177.2; ESMS m/z 349.2 [M+H]⁺; LC/MSt_(R)=8.32 (349.3 [M+H]⁺) min.

14i (W₁=5-Cl, R₁=3-methylphenyl, R₂=NH₂)

Compound 14i was synthesized using procedure (b) and obtained as agolden colored oil (40 mg, 66% yield); clogP=5.50; R_(f)(petroleumspirit/ethyl acetate, 2:1)=0.35; HPLC (214 nm) t_(R)=9.79 (92.66%) min;¹H NMR (400 MHz, CDCl₃) δ 2.35 (s, 3H), 2.40 (s, 3H), 3.67 (d, J=15.9Hz, 1H), 3.73 (d, J=15.9 Hz, 1H), 4.15 (brs, 2H), 5.38 (s, 1H), 6.61 (d,J=7.8 Hz, 1H), 7.01-7.04 (m, 2H), 7.12 (d, J=5.8 Hz, 1H), 7.24-727 (m,3H); ¹³C NMR (100 MHz, CDCl₃) δ 11.6, 21.4, 26.1, 49.8, 117.9, 123.3,125.4, 126.0, 128.5, 128.8, 128.9, 129.0, 129.6, 137.3, 138.7, 143.4,167.4, 176.9; ESMS m/z 360.1 [M+H]⁺, 401.3 [M+CH₃CN+H]⁺; LC/MSt_(R)=9.02 (360.0 [M+H]⁺) min.

14j (W₁=5-Cl, R₁=4-methylphenyl, R₂=NH₂)

Compound 14j was synthesized using procedure (b) and obtained as agolden colored oil (69 mg, 64% yield); clogP=5.50; R_(f) (petroleumspirit/ethyl acetate, 2:1)=0.41; HPLC (214 nm) t_(R)=9.90 (92.77%) min;¹H NMR (400 MHz, CDCl₃) δ 2.35 (s, 3H), 2.39 (s, 3H), 3.65 (d, J=15.9Hz, 1H), 3.72 (d, J=15.9 Hz, 1H), 3.78 (brs, 2H), 5.38 (s, 1H), 6.60 (d,J=8.2 Hz, 1H), 7.01-7.05 (m, 2H), 7.17 (d, J=7.9 Hz, 2H), 7.34 (d, J=7.9Hz, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 11.5, 21.0, 26.0, 49.6, 117.9,123.3, 125.4, 128.5, 128.8, 128.9, 129.6, 134.3, 137.9, 143.4, 167.4,176.8; ESMS m/z 360.1 [M+H]⁺, 401.2 [M+CH₃CN+H]⁺; LC/MS t_(R)=9.06(360.0 [M+H]⁺) min.

14k (W₁=5-Cl, R₁=2,3-dimethylphenyl, R₂=NH₂)

Compound 14k was synthesized using procedure (b) and obtained as agolden colored oil (20.7 mg, 30.3% yield); clogp=6.00; R_(f) (petroleumspirit/ethyl acetate, 3:1)=0.35; HPLC (214 nm) t_(R)=10.48 (95.19%) min;¹H NMR (400 MHz, CDCl₃) δ 2.03 (s, 3H), 2.28 (s, 3H), 2.40 (s, 3H), 3.68(d, J=15.9 Hz, 1H), 3.74 (d, J=15.9 Hz, 1H), 4.17 (brs, 2H), 5.63 (s,1H), 6.63 (d, J=8.4 Hz, 1H), 6.91 (d, J=2.4 Hz, 1H), 7.02 (dd, J=2.4,8.4 Hz, 1H), 7.12-7.20 (m, 2H), 7.64 (d, J=7.5 Hz, 1H); ¹³C NMR (100MHz, CDCl₃) δ 11.5, 14.4, 20.7, 26.2, 46.7, 117.7, 123.5, 125.5, 125.9,126.1, 128.4, 128.8, 129.8, 135.0, 135.6, 137.6, 143.4, 167.3, 177.0;ESMS m/z 374.2 [M+H]⁺, 367.3 [M+CH₃CN+H]⁺; LC/MS t_(R)=9.58 (244.1[M−C₄H₅N₂OS]⁺, 374.2 [M+H]⁺) min.

14l (W₁=5-Cl, R₁=3,4-dimethylphenyl, R₂=NH₂) and 14la (W₁=5-Cl,R₁=3,4-dimethylphenyl, R₂=—NHC(═O)CF₃)

Compound 14l and corresponding trifluoroacetanilide 14la weresynthesized using procedure (b). Compound 14la was obtained as a goldencolored oil (48.5 mg, 55.4% yield); clogP=6.86; R_(f) (petroleumspirit/ethyl acetate, 3:1)=0.92; HPLC (214 nm) t_(R)=10.69 (96.50%) min;¹H NMR (400 MHz, CDCl₃) δ 2.24 (s, 6H), 2.40 (s, 3H), 3.55 (d, J=15.5Hz, 1H), 3.71 (d, J=15.5 Hz, 1H), 5.59 (s, 1H), 7.07-7.14 (m, 3H), 7.32(dd, J=1.5, 8.6 Hz, 1H), 7.46 (d, J=1.5 Hz, 1H), 7.70 (d, J=8.6 Hz, 1H),9.16 (s, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 11.3, 19.3, 19.8, 25.2, 48.3,115.8 (q, J=288 Hz, CF₃), 125.6, 126.7, 128.9, 129.4, 129.7, 130.4,131.5, 133.1, 133.5, 134.5, 137.2, 137.7, 155.5 (q, J=37 Hz, COCF₃),167.4, 176.1; ESMS m/z 470.2 [M+H]⁺; LC/MS t_(R)=9.83 (469.9 [M+H]⁺)min. 14l was obtained as a golden colored oil (17.0 mg, 24.4% yield);clogP=6.00; R_(f)(petroleum spirit/ethyl acetate, 3:1)=0.34; HPLC (214nm) t_(R)=10.75 (95.51%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.25 (s, 6H),2.39 (s, 3H), 3.67 (d, J=15.9 Hz, 1H), 3.72 (d, J=15.9 Hz, 1H), 4.13(brs, 2H), 5.35 (s, 1H), 6.61 (d, J=8.4 Hz, 1H), 7.03 (dd, J=2.3, 8.5Hz, 1H), 7.07 (d, J=2.3 Hz, 1H), 7.13 (d, J=7.5 Hz, 1H) 7.18-7.21 (m,2H); ¹³C NMR (100 MHz, CDCl₃) δ 11.6, 19.4, 19.8, 26.1, 49.6, 117.8,123.3, 125.6, 126.2, 128.4, 129.0, 130.1, 130.1, 134.7, 136.5, 137.2,143.4, 167.4, 176.9; ESMS m/z 374.3 [M+H]⁺, 415.3 [M+CH₃CN+H]⁺; LC/MSt_(R)=9.40 (244.1 [M−C₄H₅N₂OS]⁺, 374.1 [M+H]⁺) min.

14m (W₁=5-Cl, R₁=2,5-dimethylphenyl, R₁=NH₂) and 14ma (W₁=5-Cl,R₁=2,5-dimethylphenyl, R₂=—NHC(═O)CF₃)

Compound 14m and 14ma were synthesized from 10m using procedure (b).14ma was obtained as a golden colored oil (19.1 mg, 40.8% yield);clogP=6.86; R_(f) (petroleum spirit/ethyl acetate, 3:1)=0.57; HPLC (214nm) t_(R)=10.72 (90.00%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.12 (s, 3H),2.35 (s, 3H), 2.39 (s, 3H), 3.60 (d, J=15.6 Hz, 1H), 3.75 (d, J=15.6 Hz,1H), 5.79 (s, 1H), 7.04-7.10 (m, 2H), 7.33-7.35 (m, 2H), 7.41 (d, J=2.1Hz, 1H), 7.74 (d, J=8.6 Hz, 1H), 8.99 (s, 1H); ¹³C NMR (100 MHz, CDCl₃)δ 11.3, 18.5, 21.1, 25.5, 45.1, 118.7 (q, J=289 Hz, CF₃), 126.5, 128.5,128.9, 129.4, 129.8, 131.4, 131.6, 133.1, 133.5, 133.7, 136.5, 155.5 (q,J=37 Hz, COCF₃), 167.4, 176.1; ESMS m/z 324.3, 470.0 [M+H]⁺; LC/MSt_(R)=9.70 (470.1 [M+H]⁺) min. 14m was obtained as a golden colored oil(5.0 mg, 13.4% yield); clogP=6.00; R_(f) (petroleum spirit/ethylacetate, 3:1)=0.24; HPLC (214 nm) t_(R)=10.27 (87.59%) min; ¹H NMR (400MHz, CDCl₃) δ 2.10 (s, 3H), 2.37 (s, 3H), 2.39 (s, 3H), 3.70 (d, J=15.8Hz, 1H), 3.75 (d, J=15.8 Hz, 1H), 4.13 (brs, 2H), 5.54 (s, 1H), 6.63 (d,J=8.6 Hz, 1H), 6.93 (s, 1H), 7.02-7.07 (m, 3H), 7.52 (s, 1H); ¹³C NMR(100 MHz, CDCl₃) δ 11.5, 18.4, 21.2, 26.3, 46.3, 117.7, 123.5, 125.2,128.5, 128.7, 131.0, 134.0, 134.9, 136.0, 143.5, 167.3, 177.0; ESMS m/z374.2 [M+H]⁺, 415.4 [M+CH₃CN+H]⁺; LC/MS t_(R)=9.60 (374.1 [M+H]⁺) min.

14o (W₁=3,5-dibromo, R₁=2-methylphenyl, R₂=NH₂)

Compound 14o was synthesized from 10o using procedure (b), isolated as aclear oil. R_(f) (EtOAc:petrol (1:10))=0.09. HPLC (214 nm) t_(R)=11.72min (98.7%). ¹H NMR (CDCl₃, 400 MHz) δ 2.11 (s, 3H), 2.42 (s, 3H), 3.71(dd, J=29, 16 Hz, 2H), 4.81 (bs, 2H), 5.61 (s, 1H), 6.95 (d, J=2 Hz,1H), 7.18 (d, J=7.2 Hz, 1H), 7.23-7.27 (m, 1H), 7.29-7.33 (m, 1H), 7.49(d, J=2 Hz, 1H), 7.78 (d, J=7.6 Hz, 1H). ESMS m/z 482.0, 484.0, 486.0[M+H]⁺, correct isotope pattern observed.

14p (W₁=5-methyl, R₁=2-methylphenyl, R₂=NH₂)

Compound 14P was synthesized from 10p using procedure (b), isolated as apale yellow oil (0.016 g, 28% yield). R_(f) (EtOAc:petrol (1:2))=0.4.HPLC (214 nm) t_(R)=8.27 min (97.1%). ¹H NMR (CDCl₃, 400 MHz) δ 2.13 (s,3H), 2.17 (s, 3H), 2.39 (s, 3H), 3.71 (dd, J=23.6, 15.6 Hz, 2H), 5.62(s, 1H), 6.61 (d, J=8 Hz, 1H), 6.75 (s, 1H), 6.87 (d, J=8 Hz, 1H),7.15-7.30 (m, 3H), 7.79 (d, J=7.2 Hz, 1H). ESMS m/z 340.4 [M+H]⁺, 381.4[M+H+ CH₃CN]⁺. LCMS (214 nm) t_(R)=7.84 min (340.0 [M+H]⁺).

14q (W, =H, R₁=2-methylphenyl, R₂=NH₂)

Compound 14o from 10q, was isolated as an orange oil. R_(f)(EtOAc:petrol (1:5))=0.09. HPLC (214 nm) t_(R)=8.06 min (5.7%), 8.81(89.9). ¹H NMR (CDCl₃, 400 MHz) δ 2.12 (s, 3H), 2.37 (s, 3H), 3.70 (dd,J=23, 16 Hz, 2H), 5.62 (s, 1H), 6.62-6.68 (m, 2H), 6.90 (dd, J=7.6, 0.8Hz, 1H), 7.02-7.06 (m, 1H), 7.14 (d, J=7.2 Hz, 1H), 7.18-7.21 (m, 1H),7.24-7.28 (m, 1H), 7.79 (d, 7.6 Hz, 1H). ESMS nL/z 326.0 [M+H]⁺, 367.4[M+H+ CH₃CN]⁺.

14t (W₁=5-nitro, R₁=phenyl, R₂=NH₂)

Compound 14d was synthesized using procedure (b) and obtained as abright yellow gum (230 mg, 0.645 mmol, 83% yield). R_(f) (silica,dichloromethane:methanol (9:1)=0.20. HPLC (214 nm) t_(R)=8.95 (93.1%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.40 (s, 3H), 3.67 (d, J=16.4 Hz, 1H),3.75 (d, J=16.4 Hz, 1H), 5.27 (br s, 2H), 5.45 (s, 1H), 6.67 (d, J=8.8Hz, 1H), 7.32-7.43 (m, 3H), 7.48-7.52 (m, 2H), 7.92 (d, J=2.4 Hz, 1H),7.97 (dd, J=8.8, 2.4 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 11.4, 25.9,49.5, 115.1, 122.2, 125.1, 126.0, 128.4, 128.8, 129.1, 136.5, 138.7,151.1, 167.2, 176.8. ESMS m/z 357.4 [(M+H)]⁺, 713.4 [(2M+H)]⁺. LC/MSt_(R)=8.23 (356.9 [(M+H)]⁺, 713.1 [(2M+H)]⁺) min.

14w (W₁=5-Cl, R₁=2,4-dimethylphenyl, R₂=NH₂) and 14wa (W₁=5-Cl,R₁=2,4-dimethylphenyl, R₂=—NHC(═O)CF₃)

Compound 14w and 14wa were synthesized using procedure (b) above. 14wawas obtained as a golden colored oil (45.7 mg, 40.8% yield); clogP=6.86;R_(f)(petroleum spirit/ethyl acetate, 3:1)=0.75; HPLC (214 nm)t_(R)=10.41 (94.58%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.14 (s, 3H), 2.31(s, 3H), 2.39 (s, 3H), 3.58 (d, J=15.6 Hz, 1H), 3.73 (d, J=15.6 Hz, 1H),5.80 (s, 1H), 7.01 (s, 1H), 7.08 (d, J=8.0 Hz, 1H), 7.33 (dd, J=2.5, 8.6Hz, 1H), 7.40 (d, J=2.5 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.73 (d, J=8.6Hz, 1H), 9.02 (s, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 11.3, 18.8, 20.9,25.5, 44.7, 126.5, 127.6, 127.9, 128.9, 129.8, 130.8, 131.6, 132.3,133.1, 133.7, 136.6, 138.5, 167.4, 176.1, Both the carbon signals forthe COCF₃ group were not observed with the number of scans used; ESMSm/z 470.2 [M+H]⁺; LC/MS t_(R)=9.85 (470.0 [M+H]⁺) min. Compound 14w wasobtained compound as a golden colored oil (18.3 mg, 20.0% yield);clogP=6.00; R_(f)(petroleum spirit/ethyl acetate, 3:1)=0.45; HPLC (214nm) t_(R)=10.60 (83.38%) min; ¹H NMR (400 MHz, CDCl₃) δ 2.11 (s, 3H),2.32 (s, 3H), 2.39 (s, 3H), 3.68 (d, J=15.8 Hz, 1H), 3.73 (d, J=15.8 Hz,1H), 4.14 (brs, 2H), 5.53 (s, 1H), 6.62 (d, J=8.5 Hz, 1H), 6.94 (d,J=2.4 Hz, 1H), 6.99 (s, 1H), 7.02 (dd, J=2.4, 8.5 Hz, 1H), 7.08 (d,J=7.9 Hz, 1H) 7.60 (d, J=7.9 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 11.5,18.8, 20.9, 26.2, 46.0, 117.7, 123.5, 125.3, 127.2, 128.2, 128.4, 128.7,131.9, 132.1, 136.9, 137.6, 143.5, 167.4, 177.0; ESMS m/z 374.3 [M+H]⁺,415.3 [M+CH₃CN+H]⁺; LC/MS t_(R)=9.74 (374.2 [M+H]⁺) min.

14y (W₁=6-CH₃, R₁=2-amino-3-pyridnyl, R₂=NH₂) was synthesized from 10vusing procedure (b), isolated as a yellow oil, which crystallised uponstanding was obtained (0.026 g, 32% yield). R_(f) (EtOAc:petrol(1:1))=0.2. HPLC (214 nm) t_(R)=6.84 min (95.9%). ¹H NMR (CDCl₃, 400MHz) δ 2.14 (s, 3H), 2.41 (s, 3H), 3.72 (dd, J=26.4, 16 Hz, 2H), 5.08(bs, 2H), 5.54 (s, 1H), 6.57 (dd, J=7.6, 4.8 Hz, 1H), 7.14-7.19 (m, 2H),7.22-7.31 (m, 2H), 7.78 (d, J=7.2 Hz, 1H), 8.00 (dd, J=5.2, 1.6 Hz, 1H).ESMS m/z 327.4 [M+H]⁺, 368.3 [M+H+ CH₃CN]⁺.

14z (W₁=5-Cl, R₁=t-butyl, R₂=NH₂) was synthesized using procedure (c)and obtained as a golden colored oil (19 mg, 14.8% yield); clogP=5.06;R_(f)(petroleum spirit/ethyl acetate, 2:1)=0.52; HPLC (214 nm)t_(R)=9.41 (82.21%) min; ¹H NMR (400 MHz, CDCl₃) δ 1.03 (s, 9H), 2.38(s, 3H), 3.34 (d, J=15.1 Hz, 1H), 3.63 (d, J=15.1 Hz, 1H), 4.19 (s, 1H),6.60 (d, J=8.4 Hz, 1H), 7.00 (dd, J=2.4, 8.4 Hz, 1H), 7.50 (d, J=2.4 Hz,1H); ESMS m/z 326.2 [M+H]⁺, 367.3 [M+CH₃CN+H]⁺; LC/MS t_(R)=8.52 (326.2[M+H]⁺) min.

14ac (W₁=H, R₁=2-methylphenyl, R₂=methoxy) was synthesized from 10acusing procedure (b) and obtained as a clear oil which crystallised uponstanding (0.029 g, 65% yield). R_(f) (EtOAc:petrol (1:10))=0.18. HPLC(214 nm) t_(R)=10.37 min (99.7%). ¹H NMR (CDCl₃, 400 MHz) δ 2.31 (s,3H), 2.34 (s, 3H), 3.72 (s, 2H), 3.33 (s, 3H), 5.98 (s, 1H), 6.83 (d,J=8.4 Hz, 1H), 6.90-6.94 (m, 1H), 7.11-7.25 (m, 4H), 7.48 (d, J=7.6 Hz,1H), 7.52 (d, J=7.6 Hz, 1H). ESMS m/z 341.3 [M+H]⁺, 681.3 [2M+H]⁺.

14ad (W₁=5-CH₃, R₁=2-methylphenyl, R₂=OH) was synthesized from 10adusing procedure (b) and obtained as a white solid (0.022 g, 40% yield).R_(f) (EtOAc:petrol (1:5))=0.15. HPLC (214 nm) t_(R)=9.30 min (99.5%).¹H NMR (CDCl₃, 400 MHz) δ 2.20 (s, 3H), 2.42 (s, 3H), 3.74 (s, 2H), 5.89(s, 1H), 6.79-6.82 (m, 1H), 6.88 (d, J=8 Hz, 1H), 7.08 (dd, J=7.6, 1.2Hz, 1H), 7.11-7.26 (m, 4H), 7.43 (bs, 1H), 7.75 (d, J=7.6 Hz, 1H). ESMSm/z 327.1 [M+H]⁺, 482.0 (unidentified)

EXAMPLE 8

15a (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2-aminoethyl)

To a stirred solution of 10s (250 mg, 0.932 mmol) in TFA (5.0 ml) undera nitrogen atmosphere at rt was added 2-aminoethanethiol hydrochloride(530 mg, 4.66 mmol, 5 eq). After stirring for 96 hrs the reactionmixture was partitioned between dichloromethane and aqueous NaOH (1mol/L). The aqueous phase was back extracted twice with dichloromethaneand the combined organic phase was dried with brine and sodium sulfate,then filtered and evaporated to give the crude thiol ether (339 mg) as abrown oil. The crude material was purified by flash chromatography onsilica (15 g) by eluting with dichloromethane:methanol (10:1) to givethiol ether 15a (222 mg, 0.678 mmol, 73% yield) as a white solid. R_(f)(silica, dichloromethane:methanol (10:1))=0.25. HPLC (214 nm) t_(R)=7.06(96.1%) min. ¹H NMR (400 MHz, CDCl₃) δ 1.00-1.70 (br s, 2H), 2.50-2.60(m, 2H), 2.80-3.00 (m, 2H), 3.85-4.40 (br s, 2H), 5.62 (s, 1H), 6.61 (d,J=8.4 Hz, 1H), 7.03 (dd, J=8.4, 2.4 Hz, 1H), 7.11 (d, J=2.4 Hz, 1H),7.23 (ddd, J=7.6, 7.6, 1.6 Hz, 1H), 7.31 (ddd, J=7.6, 7.6, 1.2 Hz, 1H),7.38 (dd, J=7.6 Hz, 1.2 Hz, 1H), 7.73 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR(100 MHz, CDCl₃) δ 36.0, 41.1, 45.4, 117.5, 123.2, 125.7, 127.2, 128.1,128.2, 128.8, 129.7, 130.5, 133.8, 136.8, 143.2. ESMS m/z 250.2[M−HS(CH₂)₂NH₂+H]⁺, 327.3 [(M+H)]⁺. LC/MS t_(R) 6.81 (250.0[M−HS(CH₂)₂NH₂+H]⁺, 327.2 [M+H]⁺, 653.3 [2M+H]⁺) min.

15b (W=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2-carboxylethyl)

Using a similar procedure as for the preparation of 15a, 15b ((R₁=5-Cl,R₂=2-chlorophenyl, R₃=2-carboxylethyl) was prepared from 10s and3-mercaptopropionic acid, obtained as a yellow gum obtained (311 mg,0.873 mmol, 47% yield). R_(f) (silica, 40:60 petroleum ether:ethylacetate (1:1)=0.42. HPLC (214 nm) t_(R)=8.28 (86.7%) min. ¹H NMR (400MHz, CDCl₃) δ 2.64-2.76 (m, 4H), 5.66 (s, 1H), 6.63 (d, J=8.4 Hz, 1H),6.83 (s, 3H), 7.03-7.08 (m, 2H), 7.25 (ddd, J=7.6, 7.6, 1.2 Hz, 1H),7.30-7.36 (m, 1H), 7.38-7.40 (m, 1H), 7.74 (dd, J=8.0, 1.2 Hz, 1H). ¹³CNMR (100 MHz, CDCl₃) δ 27.1, 34.0, 45.7, 118.1, 123.8, 125.8, 127.3,128.30, 128.34, 128.9, 129.8, 130.4, 133.9, 136.2, 142.7, 177.4. ESMSm/z 250.0 [(M−HS(CH₂)₂CO₂H+ H)]⁺, 356.1 [(M+H)]⁺. LC/MS t_(R) 8.43(250.0 [(M−HS(CH₂)₂CO₂H+ H)]⁺, 355.9 [(M+H)]⁺, 710.9 [(2M+H)]⁺) min.

15c (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=ethyl)

Using a similar procedure as for the preparation of 15a, 15c wasprepared from 10s and ethanethiol, obtained as a pale brown oil (135 mg,0.432 mmol, 47% yield). R_(f) (silica, 40-60 petroleum ether:ethylacetate (10:1)=0.20. HPLC (214 nm) t_(R)=9.97 (97.8%) min. ¹H NMR (400MHz, CDCl₃) δ 1.28 (t, J=7.2 Hz, 3H), 2.50 (m, 2H), 4.01 (br s, 2H),5.62 (s, 1H), 6.63 (d, J=8.4 Hz, 1H), 7.05 (dd, J=8.4, 2.4 Hz, 1H), 7.12(d, J=2.4 Hz, 1H), 7.25 (ddd, J=8.0, 8.0, 1.6 Hz, 1H), 7.32 (ddd, J=7.6,7.7, 1.2 Hz, 1H), 7.40 (dd, J=7.6, 1.2 Hz, 1H), 7.73 (dd, J=8.0, 1.6 Hz,1H). ¹³C NMR (100 MHz, CDCl₃) δ 14.2, 26.4, 45.2, 117.7, 123.5, 126.1,127.2, 128.0, 128.4, 128.7, 129.7, 130.5, 133.8, 136.8, 143.0. ESMS m/z250.3 [M−HSCH₂CH₃+H]⁺, 312.3 [M+H]⁺. LC/MS t_(R) 10.26 (250.0[M−HSCH₂CH₃+H]⁺, 312.0 [M+H]⁺, 622.9 [2M+H]⁺) min.

15d (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2-hydroxylethyl)

Using a similar procedure as for the preparation of 15a, 15d wasprepared from 10s and 2-mercaptoethanol, obtained as a yellow gum (142mg, 0.432 mmol, 46% yield). R_(f) (silica, 40-60 petroleum ether:ethylacetate (2:1)=0.60. HPLC (214 nm) t_(R)=7.98 (82.3%) min. ¹H NMR (400MHz, CDCl₃) δ 2.61-2.78 (m, 2H), 2.63 (t, J=5.6 Hz, 1H), 3.20-3.60 (brs, 2H), 3.78 (ddd, J=5.6, 5.6, 2.8 Hz, 2H), 5.66 (s, 1H), 6.62 (d, J=8.4Hz, 1H), 7.04 (dd, J=8.4, 2.4 Hz, 1H), 7.12 (d, J=2.4 Hz, 1H), 7.22-7.28(m, 1H), 7.30-7.36 (m, 1H), 7.39 (dd, J=8.0, 1.2 Hz, 1H), 7.72 (dd,J=8.0, 1.6 Hz, 1H). ³C NMR (100 MHz, CDCl₃) δ 35.0, 45.1, 60.7, 117.7,123.4, 125.7, 127.3, 128.2, 128.3, 128.9, 129.7, 130.3, 133.8, 136.5,143.0. ESMS m/z 328.1 [M+H]⁺, 369.3 [M+CH₃CN+H]⁺. LC/MS t_(R) 8.19(249.9 [M−HSCH₂OH+H]⁺, 328.1 [M+H]⁺, 655.2 [2M+1]⁺) min.

15e (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=(2-methoxycarbonyl)ethyl)

Using a similar procedure as for the preparation of 15a, 15e wasprepared from 10s and methyl 3-mercaptopropionate, obtained as a whitesolid (193 mg, 0.719 mmol, 38%). R_(f) (silica, 40-60 petroleumether:ethyl acetate (3:1)=0.42. HPLC (214 nm) t_(R) _(=9.13) (94.3%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.80-2.97 (m, 2H), 3.90 (s, 3H), 4.25 (brs, 2H), 5.84 (s, 1H), 6.83 (d, J=8.4 Hz, 1H), 7.22-7.29 (m, 2H),7.43-7.49 (m, 1H), 7.54 (dd, J=7.2, 7.2 Hz, 1H), 7.60 (d, J=7.2 Hz, 1H),7.92-7.97 (m, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 27.4, 33.9, 45.6, 51.8,117.7, 123.3, 125.4, 127.2, 128.2, 128.3, 128.9, 129.7, 130.3, 133.8,136.2, 143.1, 172.1. ESMS m/z 370.3 [(M+H)]⁺, 411.2 [(M+CH₃CN+H)]⁺.LC/MS t_(R) 9.51 (250.0 [(M−HS(CH₂)₂CO₂CH₃+H)]⁺, 370.0 [(M+H)]⁺,739.0[(2M+H)]⁺) min.

15ea (W₁=5-Cl, R₁=2-chlorophenyl, R₂=—NHC(═O)CF_,R₃=(2-methoxycarbonyl)ethyl)

To a stirred solution of alcohol 10s (500 mg, 1.86 mmol) in TFA (10.0ml) under a nitrogen atmosphere at rt was added methyl3-mercaptopropionate (1.03 ml, 9.32 mmol, 5 eq). After stirring for 48hrs the TFA was evaporated and the residue was partitioned betweendichloromethane and saturated aqueous sodium bicarbonate solution. Theaqueous phase was back extracted twice with further dicloromethane thenthe combined organic phase was dried with sodium sulfate, filtered andevaporated to give the crude thiol ether (1.157 g) as a brown oil. Thecrude material was purified by flash chromatography on silica (50 g) byeluting with 40-60 petroleum ether:ethyl acetate (10:1 then 5:1) to givethiol ether 15ea (510 mg, 1.09 mmol, 59% yield) as a white solid. R_(f)(silica, 40-60 petroleum ether:ethyl acetate (5:1)=0.33. HPLC (214 nm)t_(R)=9.39 (91.5%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.58-2.72 (m, 2H),3.66 (s, 3H), 5.73 (s, 1H), 7.20-7.30 (m, 2H), 7.32 (ddd, J=7.6, 7.6,1.2 Hz, 1H), 7.37 (dd, J=7.6, 1.2 Hz, 1H), 7.65 (dd, J=7.6, 1.6 Hz, 1H),7.68 (d, J=8.8 Hz), 8.97 (br s, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 27.3,30.0, 45.9, 52.0, 115.8 (q, J=287 Hz), 126.4, 127.5, 128.6, 129.2,129.5, 130.0, 130.1, 131.3, 132.7, 133.9, 135.0, 155.4 (q, J=37.2 Hz),172.2. ESMS m/z 322.9 [(unknown)]⁺, 466.3 [(M+H)]⁺, 483.1 [(M+NH₄)]⁺.LC/MS t_(R) 9.65 (345.9 [(M−HS(CH₂)₂CO₂Me⁺ H)]⁺, 466.0 [(M+H)]⁺, 931.0[(2M+H)]⁺, 948.2 [(2M+NH₄)]⁺) min.

15f (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2-pyridyl)

Using a similar procedure as for the preparation of 15a, 15f wasprepared from 10s and 2-mercaptopyridne, as a white solid (105 mg, 0.291mmol, 31% yield). R_(f) (silica, 40-60 petroleum ether:ethyl acetate(10:1)=0.25. HPLC (214 nm) t_(R)=12.03 (72.9%) min. ¹H NMR (400 MHz,CDCl₃) δ 4.31 (br s, 2H), 6.65 (d, J=8.4 Hz, 1H), 8.68 (s, 1H),6.97-7.05 (m, 2H), 7.15 (d, J=8.0 Hz, 1H), 7.24 (ddd, J=9.2, 9.2, 1.6Hz, 1H), 7.31 (ddd, J=9.6, 9.6, 1.6 Hz, 1H), 7.38 (dd, J=8.8, 1.2 Hz,1H), 7.47 (ddd, J=9.9, 9.6, 1.6 Hz, 1H), 7.85 (dd, J=9.2, 1.6 Hz, 1H),8.40-8.44 (m, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 45.7, 117.3, 120.3, 122.3,122.9, 125.7, 126.9, 128.2, 128.3, 128.8, 129.8, 130.5, 134.0, 136.4,137.2, 143.2, 149.7, 157.5. ESMS nVz 361.2 [M+H]⁺. LC/MS t_(R)=9.49(360.9 [M+H]⁺) min.

15g (W₁=5-Cl, R₁=2-chlorophenyl R₂=NH₂, R₃=4-methoxybenzyl)

Using a similar procedure as for the preparation of 15a, 15g wasprepared from 10s and 4-methoxybenzyl, obtained as a yellow oil (98 mg,0.242 mmol, 26% yield). R_(f) (silica, 40-60 petroleum ether:ethylacetate (10:1)=0.40. HPLC (214 nm) t_(R)=9.75 (81.5%) min. ¹H NMR (400MHz, CDCl₃) δ 3.46 (d, J=13.5 Hz, 1H), 3.59 (d, J=13.5 Hz, 1H), 3.63 (brs, 2H), 3.77 (s, 3H), 5.20 (s, 1H), 6.52 (d, J=8.4 Hz, 1H), 6.80-6.83(m, 2H), 6.90 (d, J=2.0 Hz, 1H), 6.97 (dd, J=8.4, 2.4 Hz, 1H), 7.05-7.10(m, 2H), 7.21-7.27 (m, 1H), 7.31-7.38 (m, 2H), 7.86 (dd, J=7.6, 1.6 Hz,1H). ¹³C NMR (100 MHz, CDCl₃) δ 36.1, 44.4, 55.2, 110.3, 114.0, 17.4,123.2, 125.4, 127.2, 128.0, 128.4, 128.8, 129.3, 129.8, 129.9, 130.7,134.2, 136.2, 142.9, 158.8. ESMS m/z 404.2 [M+H]⁺, 445.1 [M+CH₃CN+H]⁺.LC/MS t_(R) 10.69 (404.0 [M+H]⁺) min.

15h (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=n-butyl)

Using a similar procedure as for the preparation of 15a, 15h wasprepared from 10s and 1-butanethiol, obtained as a pale yellow oil (76mg, 0.223 mmol, 24% yield). R_(f) (silica, 40-60 petroleum ether:ethylacetate (10:1)=0.20. HPLC (214 nm) t_(R)=13.30 (95.2%) min. ¹H NMR (400MHz, CDCl₃) δ 0.89 (t, J=8.8 Hz, 3H), 1.35-1.46 (m, 2H), 1.54-1.64 (m,2H), 2.41-2.54 (m, 2H), 4.00 (br s, 2H), 5.58 (s, 1H), 6.63 (d, J=8.4Hz, 1H), 7.05 (dd, J=8.4, 2.4 Hz, 1H), 7.12 (d, J=2.4 Hz, 1H), 7.25(ddd, J=7.6, 7.6, 1.6 Hz, 1H), 7.33 (ddd, J=7.6, 7.6, 1.2 Hz, 1H), 7.40(dd, J=7.6, 1.2 Hz, 1H), 7.72 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz,CDCl₃) δ 13.5, 21.9, 31.1, 32.2, 45.7, 117.7, 123.6, 126.2, 127.2,128.0, 128.4, 128.7, 129.7, 130.5, 133.9, 136.9, 143.1. ESMS m/z 340.2[M+H]⁺, 381.2 [M+CH₃CN+H]⁺. LC/MS t_(R) 10.98 (249.8 [M−HS(CH₂)₃CH₃+H]⁺,340.0 [M+H]⁺) min.

15i (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2,2,2-trifluoroethyl)

Using a similar procedure as for the preparation of 15a, 15i wasprepared from 10s and 2,2,2-trifluoroethanthiol, obtained as a paleyellow oil (132 mg, 0.360 mmol, 39%). R_(f) (silica, 40-60 petroleumether:ethyl acetate (15:1)=0.33. HPLC (214 nm) t_(R)=12.13 (90.1%) min.¹H NMR (400 MHz, CDCl₃) δ 2.89-3.10 (m, 4H), 3.98 (br s, 2H), 5.85 (s,1H), 6.67 (d, J=8.4 Hz, 1H), 7.03 (d, J=2.4 Hz, 1H), 7.08 (dd, J=8.4,2.4 Hz, 1H), 7.30 (ddd, J=7.6, 7.6, 1.6 Hz, 1H), 7.34-7.40 (m, 1H), 7.44(dd, J=7.6, 1.2 Hz, 1H), 7.75 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz,CDCl₃) δ 34.1 (q, J=33.4 Hz), 46.0, 118.0, 123.6, 124.2, 127.4, 128.3,128.8, 129.4, 130.1, 134.4, 135.1, 143.3. ESMS m/z 407.1 [(M+CH₃CN+H)]⁺,LC/MS t_(R) 8.86 (365.9 [(M+H)]⁺) min.

15j (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=2-(N,N-dimethylamino)ethyl)

Using a similar procedure as for the preparation of 15a, 15l wasprepapred from 10s and 2-(dimethylamino)ethanethiol, obtained as a whitesolid (183 mg, 0.515 mmol, 55%). R_(f) (silica, 40-60 petroleumether:ethyl acetate (2:1)=0.10. HPLC (214 nm) t_(R)=7.42 (97.7%) min. ¹HNMR (400 MHz, CDCl₃) δ 2.25 (s, 6H), 2.47-2.55 (m, 4H), 4.71 (br s, 2H),5.66 (s, 1H), 6.60 (8.4 Hz, 1H), 6.67 (d, J=2.4 Hz, 1H), 6.97 (dd,J=8.4, 2.4 Hz, 1H), 7.22-7.28 (m, 1H), 7.36-7.41 (m, 2H), 8.03 (d, J=8.0Hz). ³C NMR (100 MHz, CDCl₃) δ 29.1, 45.0, 45.3, 58.4, 117.0, 122.3,125.5, 127.0, 127.7, 127.9, 128.7, 129.7, 130.8, 134.2, 136.7, 143.8.ESMS m/z 355.1 [(M+H)]⁺, LC/MS t_(R) 6.86 (354.9 [(M+H)]⁺) min.

15k (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂, R₃=3-hydroxypropyl)

To a stirred solution of ester 15e (74.0 mg, 0.199 mmol) in THF (2.5 ml)under a nitrogen atmosphere at 0° C. was added LiAlH4 (15.1 mg, 0.400mmol, 2eq). The reaction mixture was warmed to rt and then stirred for30 min. The reaction mixture was partitioned between dichloromethane (10ml) and potassium sodium tartrate (1 mol/L, 5 ml) and stirring carriedout for 1 hr at rt. Brine was added and the aqueous layer was extractedthrice with dichloromethane. The combined organic extracts were driedwith sodium sulfate, filtered and evaporated to give the crude alcohol(80.0 mg) as a brown oil. The crude material was purified by flashchromatography on silica (5 g) by eluting with 40-60 petroleumether:ethyl acetate (1:1) to give alcohol 15k (67.0 mg, 0.195 mmol, 98%yield) as a white solid. R_(f) (silica, 40-60 petroleum ether:ethylacetate (1:1))=0.40. HPLC (214 nm) t_(R)=10.56 (94.4%) min. ¹H NMR (400MHz, CDCl₃) δ 1.69 (br s, 1H), 1.78-1.87 (m, 2H). 2.49-2.63 (m, 2H),3.69 (t, J=6.0 Hz, 2H), 3.95 (br s, 2H), 5.57 (s, 1H), 6.61 (d, J=8.8Hz, 1H), 7.03 (dd, J=8.8, 2.4 Hz, 1H), 7.14 (d, J=2.4 Hz, 1H), 7.19-7.26(m, 1H), 7.27-7.33 (m, 1H), 7.37 (dd, J=8.0, 1.2 Hz, 1H), 7.66 (dd,J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) 829.1, 31.7, 45.6, 61.4,117.8, 123.6, 126.0, 127.3, 128.2, 128.5, 128.8, 129.7, 130.4, 133.8,136.7, 143.0. ESMS m/z 342.2 [M+H]⁺, 383.2 [M+CH₃CN+H]⁺. LC/MSt_(R)=8.34 (249.8 [M−HS(CH₂)₃OH+H]⁺, 341.9 [M+H]⁺, 682.9 [2M+H]⁺) min.

15l (W₁=5-Cl, R₁=cyclohexyl, R₂=NHBoc, R₃=2-aminoethyl

EXAMPLE 9

Synthesis of 16a (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=phenyl)

To a stirred solution of primary amine 15a (100 mg, 0.305 mmol) in THF(2.5 ml) under a nitrogen atmosphere at rt was added benzoic acid (37.3mg, 0.305 mmol, 1 eq), EDC (70.3 mg, 0.367 mmol, 1.2eq) and DMAP (3.7mg, 0.030 mmol, 0.1 eq.). After stirring for 18 hrs the reaction mixturewas partitioned between dichloromethane and brine. The aqueous phase wasback extracted twice with dichloromethane and the combined organic phasewas dried with sodium sulfate, then filtered and evaporated to give thecrude amide (167 mg) as a brown oil. The crude material was purified byflash chromatography first on silica (5 g) by eluting with 40-60petroleum ether:ethyl acetate (2:1) to give amide 16a (108 mg, 0.250mmol, 82% yield) as a white solid. R_(f) (silica, 40-60 petroleumether:ethyl acetate (2:1))=0.20. HPLC (214 nm) t_(R)=9.93 (99.1%) min.¹H NMR (400 MHz, CDCl₃) δ2.65-2.74 (m, 2H), 3.46-3.59 (m, 1H), 3.64-3.75(m, 1H), 3.96 (br s, 2H), 5.62 (s, 1H), 6.58 (d, J=8.4 Hz, 1H), 6.76(dd, J=5.2 Hz, 1H), 7.02 (dd, J=8.4, 2.4 Hz, 1H), 7.18-7.29 (m, 3H),7.34-7.42 (m, 3H), 7.44-7.50 (m, 1H), 7.60 (dd, J=7.6, 1.6 Hz, 1H),7.39-7.77 (m, 2H). ³C NMR (100 MHz, CDCl₃) δ 32.5, 38.2, 44.8, 117.8,123.3, 125.2, 126.8, 127.3, 128.2, 128.4, 128.5, 128.9, 129.7, 130.1,131.4, 133.7, 134.1, 136.3, 143.0, 167.5. ESMS m/z 247.3 [unknown]⁺,430.9 [(M+H)]⁺. LC/MS t_(R)=9.11 (431.1 [(M+H)]⁺) mm.

Using a similar procedure as for the synthesis of 16a, except thesolvent was changed from THF to DCM, the following amides were prepared.

16b (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=3,4-dimethoxybenzyl)

Form 15a and (3,4-dimethoxyphenyl)acetic acid, compound 16b was obtainedas a white solid (110 mg, 0.217 mmol, 89% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.20. HPLC (214 nm) t_(R)=9.33 (97.9%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.48-2.62 (m, 2H), 3.21-3.31 (m, 1H),3.46-3.57 (m, 3H), 3.80 (s, 3H), 3.86 (s, 3H), 4.06 (br s, 2H), 5.54 (s,1H), 5.91 (t, J=1.6 Hz, 1H), 6.59 (d, J=8.4 Hz, 1H), 6.76 (m, 3H), 7.02(dd, J=8.4, 4.2 Hz, 1H), 7.13 (d, J=1.6 Hz, 1H), 7.19-7.30 (m, 2H), 7.36(dd, J=8.4, 1.6 Hz, 1H), 7.58 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz,CDCl₃) δ 32.3, 37.7, 43.1, 44.5, 55.7, 55.8, 111.3, 112.3, 117.7, 121.6,123.2, 125.1, 126.9, 127.3, 128.2, 128.3, 128.9, 129.7, 130.1, 133.7,136.2, 143.1, 148.1, 149.1, 171.4. ESMS m/z 505.3 [(M+H)]⁺. LC/MSt_(R)=8.64 (505.2 [(M+H)]⁺) min.

16c (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=2,4-dichlorophenyl)

From 15a and 2,4-dichlorobenzoic acid, compound 16c was obtained as awhite solid (89.0 mg, 0.177 mmol, 83% yield). R_(f) (silica,dichloromethane:methanol (100:1))=0.20. HPLC (214 nm) t_(R)=10.65(96.1%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.64-2.79 (m, 2H), 3.48-3.58 (m,1H), 3.65-3.83 (m, 3H), 5.62 (s, 1H), 6.59 (d, J=8.4 Hz, 1H), 6.64-6.72(m, 1H), 7.19-7.30 (m, 4H), 7.34-7.40 (m, 2H), 7.55 (d, J=8.4 Hz, 1H),7.60 (dd, J=7.6, 1.6 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 32.4, 38.6,45.1, 117.9, 123.5, 125.4, 127.4, 128.3, 128.5, 129.0, 129.8, 129.9,130.2, 131.1, 131.5, 133.1, 133.7, 136.4, 136.7, 143.0, 165.5. ESMS m/z499.2 [(M+H)]⁺. LC/MS t_(R)=9.83 (498.8 [(M+H)]⁺) min.

16d (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=3,4-dimethoxyphenyl)

From 15a and 3,4-dimethoxybenzoic acid, compound 16d was obtained as awhite solid (70.0 mg, 0.142 mmol, 66% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.20. HPLC (214 nm) t_(R)=9.54 (99.5%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.68-2.79 (m, 2H), 3.49-3.59 (m, 1H),3.65-3.75 (m, 1H), 3.89 (s, 3H), 3.90 (s, 3H), 3.85-4.05 (br s obs, 2H),5.63 (s, 1H), 6.57-6.63 (m, 2H), 6.82-6.87 (m, 1H), 7.01-7.06 (m, 1H),7.19-7.32 (m, 4H), 7.35-7.43 (m, 2H), 7.57-7.61 (m, 1H). ¹³C NMR (100MHz, CDCl₃) δ 32.6, 38.3, 44.9, 55.9, 110.3, 110.6, 117.9, 119.4, 123.5,125.4, 126.9, 127.4, 128.3, 128.5, 129.0, 129.8, 130.2, 133.7, 136.5,143.1, 148.9, 151.8, 167.0. ESMS m/z 491.2 [(M+H)]⁺. LC/MS t_(R)=8.86(491.0 [(M+H)]⁺, 980.9 [(2M+H)]⁺) min.

16e (W₁=5Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=3-bromophenyl)

From 15a and 3-bromobenzoic acid, compound 16e was obtained as a whitesolid (93.0 mg, 0.182 mmol, 85% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.20. HPLC (214 nm) t_(R)=10.64 (96.1%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.65-2.79 (m, 2H), 3.49-3.59 (m, 1H),3.63-3.73 (m, 1H), 3.86 (br s, 2H), 5.61 (s, 1H), 6.59 (d, J=8.4 Hz,1H), 6.61-6.70 (m, 1H), 7.03 (dd, J=8.4, 3.6 Hz, 1H), 7.19-7.30 (m, 4H),7.37 (dd, J=7.6, 1.2 Hz, 1H), 7.56-7.62 (m, 2H), 7.65 (dd, J=7.6, 1.6Hz, 1H), 7.89 (d, J=1.6 Hz, 1H). ³C NMR (100 MHz, CDCl₃) δ 32.5, 38.5,45.0, 117.9, 122.7, 123.6, 125.4, 127.4, 128.4, 128.5, 129.0, 129.8,130.0, 130.2, 133.7, 134.4, 136.2, 136.4, 143.0, 166.1. ESMS m/z 509.1[(M+H)]⁺. LC/MS t_(R)=9.79 (509.1 [(M+H)]⁺) min.

16f (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=4-methoxycyclohexyl).

From 15a and 4-methoxycyclohexanecarboxylic acid, compound 16f wasobtained as a yellow oil (89.0 mg, 0.190 mmol, 89% yield). R_(f)(silica, dichloromethane:methanol (100:1))=0.20. HPLC (214 nm)t_(R)=9.29 (42.6%) and 9.46 (56.6%) min. (This compound is a mixture ofcis and trans isomers due to the nature of the reagent used. The totalpurity of this mixture is 99.2%.) ¹H NMR (400 MHz, CDCl₃) δ 1.12-1.27(m, 1H), 1.37-1.56 (m, 2H), 1.60-1.67 (m, 1H), 1.70-1.82 (m, 1H),1.86-2.05 (m, 2H), 2.08-2.17 (m, 1H), 2.52-2.66 (m, 2H), 3.08-3.16 (m,1H), 3.29 (s, 3H), 3.30 (s obs, 0.5H), 3.33 (s, 3H), 3.40-3.45 (m,0.5H), 3.46-3.57 (m, 1H), 4.10 (br s, 2H), 5.59 (s, 1H), 5.95-6.00 (m,1H), 6.60 (d, J=8.4 Hz, 1H), 7.00-7.04 (m, 1H), 7.17 (d, J=2.4 Hz, 1H),7.20-7.32 (m, 3H), 7.38 (d, J=8.0 Hz, 1H), 7.59 (d, J=8.0 Hz, 0.5H),7.63 (d, J=7.6 Hz, 0.5 Hz). ³C NMR (100 MHz, CDCl₃) δ 23.8, 27.7, 28.6,30.9, 32.7, 37.5, 37.6, 44.3, 44.5, 44.8, 55.4, 55.5, 74.1, 78.3, 117.8,123.2, 125.3, 127.3, 128.2, 128.4, 128.9, 129.8, 130.2, 133.8, 136.5,143.1, 175.5, 175.6. ESMS m/z 467.1 [(M+H)]⁺. LC/MS t_(R)=8.55 (467.0[(M+H)]⁺, 933.3 [(2M+H)]⁺) and 8.73 (467.0 [(M+H)]⁺, 933.0 [(2M+H)]⁺)min.

16g (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=4-nitrophenyl)

From 15a and 4-nitrobenzoic acid, compound 16g was obtained as a yellowgum (100 mg, 0.209 mmol, 98% yield). R_(f) (silica,dichloromethane:methanol (100:1))=0.20. HPLC (214 nm) t_(R)=10.19(94.1%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.75-2.80 (m, 2H), 3.50 (br s,2H), 3.52-3.68 (m, 1H), 3.68-3.81 (m, 1H), 5.62 (s, 1H), 6.62 (d, J=8.4Hz, 1H), 6.69 (br s, 1H), 7.06 (dd, J=8.4, 2.4 Hz, 1H), 7.21-7.31 (m,2H), 7.35-7.41 (m, 1H), 7.50-7.55 (m, 1H), 7.92 (d, J=8.8 Hz, 2H), 8.28(d, J=8.8 Hz, 2H). ³C NMR (100 MHz, CDCl₃) δ 32.5, 38.6, 44.9, 118.1,123.8, 125.5, 127.6, 127.8, 128.1, 128.6, 129.2, 129.9, 130.1, 133.6,136.4, 139.8, 142.8, 149.6, 165.5. ESMS m/z 476.2 [(M+H)]⁺. LC/MSt_(R)=9.40 (475.9 [(M+H)]⁺) min.

16h (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=cyclobutyl)

From 15a and cyclobutanecarboxylic acid, compound 16h was obtained as awhite solid (75.0 mg, 0.183 mmol, 85% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.20. HPLC (214 nm) t_(R)=9.50 (99.1%)min. ¹H NMR (400 MHz, CDCl₃) δ 1.80-2.00 (m, 2H), 2.08-2.18 (m, 2H),2.20-2.30 (m, 2H), 2.53-2.67 (m, 2H), 2.92-3.02 (m, 1H), 3.29-3.38 (m,1H), 3.47-3.57 (m, 1H), 3.90 (br s, 2H), 5.59 (s, 1H), 5.76 (br s, IfH),5.61 (d, J=8.4 Hz, 1H), 7.03 (dd, J=8.4, 2.0 Hz, 1H), 7.19-7.32 (m, 3H),7.39 (dd, J=7.6, 1.6 Hz, 1H), 7.60 (dd, J=7.6, 1.6 Hz 1H). ¹³C NMR (100MHz, CDCl₃) δ 18.1, 25.3, 32.7, 37.7, 39.8, 44.9, 117.8, 123.4, 125.4,127.4, 128.3, 128.5, 129.0, 129.8, 130.2, 133.8, 136.5, 143.1, 175.0.ESMS m/z 409.2 [(M+H)]⁺. LC/MS t_(R)=8.77 (409.2 [(M+H)]⁺, 817.2[(M+H)]⁺) min.

16i (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=cyclohexyl)

From 15a and cyclohexanecarboxylic acid, compound 16i was obtained as awhite solid (94.0 mg, 0.214 mmol, 100% yield). R_(f) (silica,dichloromethane:methanol (100:1))=0.20. HPLC (214 nm) t_(R)=10.23(95.7%) min. ¹H NMR (400 MHz, CDCl₃) δ 1.16-1.34 (m, 3H), 1.34-1.48 (m,2H), 1.62-1.71 (m, 1H), 1.71-1.98 (m, 3H), 2.00-2.12 (m, 1H), 2.54-2.68(m, 2H), 3.28-3.38 (m, 1H), 3.47-3.58 m, 1H), 3.97 (br s, 2H), 5.94 (s,1H), 5.84 (br s, 1H), 6.61 (d, J=8.4 Hz, 1H), 7.03 (dd, J=8.4, 2.4 Hz,7.20-7.32 (m, 3H), 7.37-7.42 (m, 1H), 7.58-7.62 (m, 1H). ¹³C NMR (100MHz, CDCl₃) δ 25.6, 28.8, 29.6, 32.8, 37.6, 44.9, 45.4, 117.9, 123.5,125.4, 127.4, 128.3, 128.5, 129.0, 129.8, 130.2, 133.8, 136.6, 143.1,17.2. ESMS fnlz 137.1 [(M+H)]⁺. LC/MS t_(R)=9.42 (437.1 [(M+H)]⁺, 873.1[(2M+H)]⁺) min.

16j (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=isopropyl)

From 15a and isobutyric acid, compound 16j was obtained as a colourlessgum (74.0 mg, 0.186 mmol, 87% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.20. HPLC (214 nm) t_(R)=9.30 (98.8%)min. ¹H NMR (400 MHz, CDCl₃) δ 1.14 (d, J=6.8 Hz, 6H), 2.34 (septet,J=6.8 Hz, 1H), 2.54-2.68 (m, 2H), 3.28-3.38 (m, 1H), 3.47-3.57 (m, 1H),4.05 (br s, 2H), 5.60 (s, 1H), 5.91 (br s, IfH), 6.61 (d, J=8.4 Hz, 1H),7.03 (dd, J=8.4, 1.6 Hz, 1H), 7.19-7.32 (m, 3H), 7.38 (d, J=7.6 Hz, 1H),7.61 (d, J=7.6 Hz, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 19.5, 32.7, 35.5,37.6, 44.9, 117.8, 123.4, 125.3, 127.3, 128.3, 128.5, 128.9, 129.8,130.2, 133.8, 136.5, 143.1, 177.1. ESMS m/z 397.3 [(M+H)]⁺. LC/MSt_(R)=8.57 (397.0 [(M+H)]⁺, 792.9 [(2M+H)]⁺) min.

16k (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=3-pyridyl)

From 15a and nicotinic acid, compound 16k was obtained as a white solid(77.0 mg, 0.178 mmol, 83% yield). R_(f) (silica,dichloromethane:methanol (40:1))=0.10. HPLC (214 nm) t_(R)=7.80 (98.4%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.72-2.79 (m, 2H), 3.2-4.0 (m, 4H), 5.62(s, 1H), 6.60 (d, J=8.4 Hz, 1H), 6.77-6.88 (m, 1H), 7.03 (dd, J=8.4, 2.4Hz, 1H), 7.19-7.30 (m, 3H), 7.30-7.40 (m, 2H), 7.57 (dd, J=7.6, 1.6 Hz,1H), 8.07-8.11 (m, 1H), 8.70 (br s, 1H), 8.97 (br s, 1H). ¹³C NMR (100MHz, CDCl₃) δ 32.5, 38.5, 45.0, 118.0, 123.4, 123.6, 125.4, 127.5,128.4, 128.5, 129.1, 129.8, 130.2, 133.7, 135.0, 136.4, 143.0, 147.9,152.2, 165.6. ESMS m/z 432.1 [(M+H)]⁺, 473.3 [(M+CH₃CN+H)]⁺. LC/MSt_(R)=7.17 (432.1 [(M+H)]⁺, 863.0 [(2M+1)]⁺) min.

16l (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H=R_(3d)=phenoxymethyl)

From 15a and phenoxyacetic acid, compound 16l was obtained as a palebrown gum (101 mg, 0.218 mmol, 102% yield). R_(f) (silica,dichloromethane:methanol (100:1))=0.20. HPLC (214 nm) t_(R)=10.26(98.4%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.66 (br s, 2H), 3.41-3.48 (m,1H), 3.61-3.68 (m, 1H), 4.05 (br s, 2H), 4.50 (s, 2H), 5.65 (br s, 1H),6.59 (br s, 1H), 6.90-7.09 (m, 5H), 7.15-7.20 (m, 1H), 7.20-7.36 (m,4H), 7.39 (dd, J=7.6, 1.2 Hz, 1H), 7.64 (dd, J=7.6, 1.2 Hz, 1H). ³C NMR(100 MHz, CDCl₃) δ 32.3, 37.6, 45.0, 67.3, 114.7, 118.2, 122.2, 123.9,125.6, 127.4, 128.3, 129.0, 129.8, 129.9, 130.3, 133.9, 136.4, 157.1,168.6. ESMS m/z 461.1 [(M+H)]⁺. LC/MS t_(R)=9.49 (461.0 [(M+H)]⁺) min.

16m (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=2-pyridyl)

From 15a and picolinic acid, compound 16m was obtained as a white solid(70.0 mg, 0.162 mmol, 75% yield). R_(f) (silica,dichloromethane:methanol (100:1))=0.20. HPLC (214 nm) t_(R)=9.73 (96.7%)min. ¹H NMR (400 MHz, CDCl₃) δ 2.70-2.77 (m, 2H), 3.56-3.63 (m, 3H),3.72-3.78 (m, 1H), 5.69 (s, 1H), 6.60 (d, J=8.4 Hz, 1H), 7.11 (d, J=2.8Hz, 1H), 7.20-7.33 (m, 2H), 7.35-7.45 (m, 2H), 7.72 (dd, J=7.6, 1.6 Hz,1H), 7.84 (ddd, J=7.6, 1.6, 1.6 Hz, 1H), 8.17 (d, J=7.6 Hz, 1H),8.28-8.39 (m, 1H), 8.54-8.57 (m, 1H). ¹³C NMR (100 MHz, CDCl₃) δ 32.5,38.2, 45.2, 117.8, 122.2, 123.4, 125.5, 126.2, 127.3, 128.3, 128.5,128.9, 129.8, 130.4, 134.0, 136.5, 137.3, 143.2, 148.1, 149.6, 164.4.ESMS m/z 432.0 [(M+H)]⁺. LC/MS t_(R)=8.93 (432.1 [(M+H)]⁺, 863.1[(2M+H)]⁺) min.

Synthesis of 16n (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H,R_(3d)=3,4-dimethoxyphenyl)

To a stirred solution of compound 15l (0.48 g, 1.34 mmol) in dry CH₂Cl₂(10 mL), 3,4-dimethoxybenzoic acid (0.25 g, 1.34 mmol), EDC (0.31 g, 1.6mmol) and DMAP (3 mg, 0.27 mmol) were added and the solution was stirredat room temperature for 24 hrs. The reaction was diluted with additionalCH₂Cl₂ (20 mL) and washed with a saturated solution of NaHCO₃ (2×10 mL),water (10 mL) and brine (20 mL). The organic layer was dried over sodiumsulfate, filtered and concentrated under vacuum. The crude product waspurified by flash chromatography on a silica gel column using a mixtureof petroleum ether and ethyl acetate as eluent to give 16n assemi-transparent solid (0.5 g, 83%): LC/MS calcd for C₂₄H₃₁ClN₂O₃S: 462[M+Na]⁺, found: 486.

16o (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=3,4,5-trimethoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16o was preparedfrom 15l and 3,4,5-trimethoxybenzoic acid as a semi-transparent solid(90% yield): LC/MS calcd for C₂₅H₃₃ClN₂O₄S: 492 [M+Na⁺], found: 515.

16p (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H,R_(3d)=3,5-dimethoxy-4-hydroxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16p was preparedfrom 15l and 3,5-dimethoxy-4-hydroxybenzoic acid as a semi-transparentsolid (88% yield): LC/MS calcd for C₂₄H₃₁ClN₂O₄S: 478 [M+Na⁺], found:501.

16q (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=2-methoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16q was preparedfrom 15l and o-Anisic acid as a semi-transparent solid (77% yield):LC/MS calcd for C₂₃H₂₉Cl N₂O₂S: 432 [M+Na⁺], found: 455).

16r (W₁=5Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=3-methoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16r was preparedfrom 15l and m-Anisic acid as a semi-transparent solid (83% yield):LC/MS calcd for C₂₃H₂₉Cl N₂O₂S: 432 [M+Na⁺], found: 455.

16s (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=4-methoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16s was preparedfrom 15l and p-Anisic acid as a semi-transparent solid (86% yield):LC/MS calcd for C₂₃H₂₉Cl N₂O₂S: 432 [M+Na⁺], found: 455.

16t (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=3-pyridyl)

Using a similar procedure as for the synthesis of 16n, 16t was preparedfrom 15l and nicotinic acid as a semi-transparent solid (75% yield):LC/MS calcd for C₂₁H₂₆ClN₃OS: 403 [M+Na⁺], found: 425.

16u (W₁=5Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=4-pyridyl)

Using a similar procedure as for the synthesis of 16n, 16u was preparedfrom 15l and isonicotinic acid as a semi-transparent solid (83% yield):LC/MS calcd for C₂₁H₂₆Cl N₃OS: 403 [M−C₈H₉N₂OS)], found: 222.

16v (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=4-pyrazolyl)

Using a similar procedure as for the synthesis of 16n, 16v was preparedfrom 15l and 4-pyrazolcarboxylic acid as a semi-transparent solid (86%yield): LC/MS calcd for C₁₉H₂₅ClN₄OS: 392 [M+Na⁺], found: 415.

16w (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=piperonyl)

Using a similar procedure as for the synthesis of 16n, 16w was preparedfrom 15l and piperonylic acid as a semi-transparent solid (88% yield):LC/MS calcd for C₂₃H₂₇ClN₂O₃S: 446 [M+Na⁺], found: 469.

16x (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3d)=4-(methylsulfonyl)phenyl

Using a similar procedure as for the synthesis of 16n, 16x was preparedfrom 15l and 4-(methylsulfonyl) benzoic acid as a semi-transparent solid(76% yield): LC/MS calcd for C₂₃H₂₉ClN₂O₃S₂: 480 [M+Na⁺], found: 503.

16y (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3b)=—(C₆H₅)SO₂NH₂)

Using a similar procedure as for the synthesis of 16n, 16v was preparedfrom 15l and 4-carboxybenzene sulfonamide as a semi-transparent solid(84% yield): LC/MS calcd for C₂₂H₂₈ClN₃O₃S₂: 481 [M+Na⁺], found: 504.

16z (W₁=5-Cl, R₁=iso-propyl, R₄/R₄=H, R_(3d)=3,4-dimethoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16z was preparedfrom an analog of 15l and 3,4-dimethoxybenzoic acid as asemi-transparent solid (90% yield): LC/MS calcd for C₂₁H₂₇ClN₂O₃S: 422[M+Na⁺], found: 445.

16aa (W₁=5-Cl, R₁=tert-butyl, R₄/R₄=H, R_(3d)=3,4-dimethoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16aa was preparedfrom an analog of 15l and 3,4-dimethoxybenzoic acid as asemi-transparent solid (85% yield): LC/MS calcd for C₂₂H₂₉ClN₂O₃S: 436[M+Na⁺], found: 459.

16ab (W₁=5-Cl, R₁=iso-propyl, R₄/R₄=H, R_(3d)=3,4,5-trimethoxyphenl

Using a similar procedure as for the synthesis of 16n, 16ab was preparedfrom an analog of 15l and 3,4,5-trimethoxybenzoic acid as asemi-transparent solid (85% yield): LC/MS calcd for C₂₂H₂₉ClN₂O₄S: 452[M+Na⁺], found: 475.

16ac (W₁=5-Cl, R₁=tert-butyl, R₄/R₄=H, R_(3d)=3,4,5-trimethoxyphenyl)

Using a similar procedure as for the synthesis of 16n, 16ac was preparedfrom an analog of 15l and 3,4,5-trimethoxybenzoic acid as asemi-transparent solid (80% yield): LC/MS calcd for C₂₃H₃₁ClN₂O₄S: 466[M+Na⁺], found: 489.

Alternate Synthesis of 16n (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H,R_(3d)=3,4-dimethoxyphenyl)

To a solution of 10ak (0.46 g, 1.36 mmol) in dry CH₂Cl₂ (5 mL), A (0.4g, 1.63 mmol) and TFA (5 mL) were added and stirred for 12 hrs. The TFAwas pumped off and the residue was dissolved in CH₂Cl₂ (30 mL) andwashed with saturated solution of NaHCO₃ (2x10 mL), water (10 mL) andbrine (20 mL). The organic layer was dried over sodium sulfate, filteredand concentrated under vacuum. The crude product was purified by flashchromatography on a silica gel column using a mixture of petroleum etherand ethyl acetate as eluent to give 16n as a semi-transparent solid(0.58 g, 92% yield): LC/MS calcd for C₂₄H₃₁ClN₂O₃S: 462 [M+Na⁺], found:486.

Alternate Synthesis of 16o (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H,R_(3d)=3,4,5-trimethoxyphenyl)

Using a similar procedure as for the alternate synthesis of 16n, 16o wasprepared from 10ak and B as a semi-transparent solid (91% yield): LC/MScalcd for C₂₅H₃₃ClN₂O₄S: 492 [M+Na⁺], found: 515.

EXAMPLE 10

17a (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3a)=3-fluorophenyl)

Amine 15a (10 mg, 0.031 mmol) was dissolved in dichloromethane (0.5 ml)and 3-fluorophenylisocyanate (3 μL, 0.031 mmol, 1 eq) was added. Afterstanding for 20 hrs at rt, the solvent was removed. The crude materialwas purified by filtration through a plug of silica (EtOAc/LP 1:1, v/v)and evaporated to dryness to yield 17a as a white solid (11 mg, 0.023mmol, 77%). R_(f) (silica, EtOAc/LP (1:2))=0.21. HPLC (214 nm)t_(R)=10.16 (>98%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.55-2.65 (m, 2H),2.70-3.30 (br s, 2H), 3.30-3.40 (m, 1H), 3.45-3.55 (m, 1H), 5.30-5.40(m, 1H), 5.60 (s, 1H), 6.59 (d, J=8.4 Hz, 1H), 6.70-6.76 (m, 1H), 6.88(s, 1H), 6.92-7.00 (m, 1H), 7.03 (dd, J=8.4, 2.4 Hz, 1H), 7.14-7.32 (m,4H), 7.36 (dd, J=7.8, 1.2 Hz, 1H), 7.59 (dd, J=7.6, 1.6 Hz, 1H). ESMSm/z 464.0 [(M+H)]⁺. LC/MS t_(R) 9.93 (464.0 [(M+H)]⁺) min.

Using similar procedure as for the synthesis of 17a, the followingthioether ureas were synthesized.

17b (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H,R_(3a)=2-trifluoromethoxyphenyl, R_(3b)=H)

From 15a and 2-trifluoromethoxyphenyl isocyanate, compound 17b wasobtained as a white solid (58.9 mg, 73% yield). LC-MS: calcd. forC₂₃H₂₀Cl₂F₃N₃O₂S: 529.06; found: 551.9 [M+Na]⁺.

17c (W₁5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3a)=3,5-dichlorophenyl,R_(3b)=H)

From 15a and 3,5-dichlorophenyl isocyanate, compound 17c was obtained asa white solid (59.5 mg, 76% yield). LC-MS: calcd. for C₂₂H₁₉Cl₄N₃OS:513.00; found: 513.8 [M+H]⁺.

17d (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3a)=2,5-difluorophenyl,R_(3b)=H)

From 15a and 2,5-difluorophenyl isocyanate, compound 17d was obtained asa white solid (53.9 mg, 74% yield). LC-MS: calcd. for C₂₂H₁₉Cl₂F₂N₃OS:481.06; found: 481.8 [M+H]⁺.

17e (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3a)=23-dichlorophenyl,R_(3b) =H)

From 15a and 2,3-dichlorophenyl isocyanate, compound 17e was obtained asa white solid (53.9 mg, 74% yield). LC-MS: calcd. for C₂₂H₁₉Cl₄N₃OS:513.00; found: 513.8 [M+H]⁺.

17f (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3a)=2,4-dimethoxyphenyl,R_(3b)=H)

From 15a and 2,4-dimethoxyphenyl isocyanate, compound 17f was obtainedas a colorless gum (41.9 mg, 74% yield). LC-MS: calcd. forC₂₄H₂₅Cl₂N₃O₃S: 505.10 found: 528.0 [M+Na]⁺.

Synthesis of 17g (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H,R_(3a)=2,4-dimethoxyphenyl, R_(3b)=H)

To a solution of 15l (0.070 g, 0.23 mmol) in dry CH₂Cl₂ (5 mL),2,4-dimethoxyphenyl isocyanate (0.046 g, 2.6 mmol) in CH₂Cl₂ (2 mL) wasadded over a period of 8 hrs and stirred additionally for 16 hrs at roomtemperature. The reaction was diluted with additional CH₂Cl₂ (20 mL) andwashed with a saturated solution of NaHCO₃ (2×10 mL), water (10 mL) andbrine (20 mL). The organic layer was dried over sodium sulfate, filteredand concentrated under vacuum. The crude product was purified by flashchromatography on a silica gel column using a mixture of petroleum etherand ethyl acetate as eluent to give 17g as a semi-transparent solid(0.08 g, 73% yield): LC/MS calcd for C₂₄H₃₂ClN₃O₃S: 477 [M+Na⁺], found:500.

17h (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3a)=3,4,5-trimethoxyphenyl,R_(3b)=H

Using a similar procedure as for the synthesis of 17g, 17h was preparedfrom 15l and 3,4,5-trimethoxyphenyl isocyanate as a semi-transparentsolid (88% yield): LC/MS calcd for C₂₅H₃₄ClN₃O₄S: 507 [M+Na⁺], found:530.

Synthesis of 17i

Using a similar procedure as for the synthesis of 17w, 17i was preparedfrom 15l and 3,4,5-trimethoxyphenyl isothiocyanate as a semi-transparentsolid (62% yield): LC/MS calcd for C₂₅H₃₄ClN₃O₃S₂: 523 [M+Na⁺], found:546.

17i (W₁=5-Cl, R₁=cyclohexyl, R₄/R₄=H, R_(3a)=3,4-dimethoxyphenyl,R_(3b)=H)

Using a similar procedure as for the synthesis of 17R, 17i was preparedfrom 15l and 3,4-dimethoxyphenyl isocyanate as a semi-transparent solid(83% yield): LC/MS calcd for C₂₄H₃₂ClN₃O₃S: 477 [M+Na⁺], found: 500.

Synthesis of 17k

Using a similar procedure as for the synthesis of 172, 17k was preparedfrom 15l and 2,4-dimethoxyphenyl isothiocyanate as a semi-transparentsolid (70% yield): LC/MS (C₂₄H₃₂ClN₃O₂S₂: 493 [M+Na⁺], found: 516.

EXAMPLE 11

18a (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=3-trifluoromethylphenyl

Amine 15a (10 mg, 0.031 mmol) was dissolved in dichloromethane (0.5 ml)and 3-trifluorobenzenesulfonyl chloride (5 μL, 0.031 mmol, 1 eq) andDIEA (11 μL, 0.062 mmol, 2 eq) were added. After standing for 20 hrs atrt, the solvent was removed. The crude material was purified byfiltration through a plug of silica (EtOAc/LP 1:1, v/v) and evaporatedto dryness to yield 18a (12 mg, 0.022 mmol, 73%) as a white solid. R_(f)(silica, EtOAc/LP (1:2))=0.40. HPLC (214 nm) t_(R)=10.84 (92.7%) min. ¹HNMR (400 MHz, CDCl₃) δ 2.55-2.67 (m, 2H), 2.75-3.09 (br s, 2H),3.09-3.25 (m, 2H), 5.21 (t, J=5.8 Hz, 1H), 5.50 (s, 1H), 6.62 (d, J=8.4Hz, 1H), 7.04 (dd, J=8.4, 2.4 Hz, 1H), 7.21-7.29 (m, 3H), 7.37 (dd,J=7.6, 1.6 Hz, 1H), 7.51 (dd, J=7.6, 2.0 Hz, 1H), 7.65 (t, J=7.8 Hz,1H), 7.83 (d, J=7.6 Hz, 1H), 8.02 (d, J=8.0 Hz, 1H), 8.11 (s, 1H). ESMSm/z 535.1 [(M+H)]⁺, 576.2 [(M+CH₃CN+H)]⁺. LC/MS t_(R) 10.65 (315.1[(unknown)]⁺, 535.0 [(M+H)]⁺) min.

Using similar procedure as for the synthesis of 18a, the followingthioether sulfonamides were synthesized.

18b (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=phenyl)

From 15a and benzenesulfonyl chloride, compound 18b was obtained as awhite solid (50.7 mg, 71%). LC-MS: calcd. for C₂₁H₂₀Cl₂N₂O₂S₂: 466.03found: 466.8 [M+H]⁺.

18c (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=2-nitrophenyl)

From 15a and 2-nitrobenzenesulfonyl chloride, compound 18c was obtainedas a white solid. LC-MS: calcd. for C₂₁H₁₉Cl₂N₃O₄S₂: 511.02 found: 533.9[M+H]⁺.

18d (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H,R_(3d)=4-trifluoromethoxyphenyl)

From 15a and 4-trifluoromethoxybenzenesulfonyl chloride, compound 18dwas obtained as a white solid. LC-MS: calcd. for C₂₂H₁₉Cl₂F₃N₂O₃S₂:550.02 found: 550.8 [M+H]⁺.

18e (W₁=5-Cl, R₁=2-chlorophenyl, R₄/R₄=H, R_(3d)=methyl)

From 15a and methanesulfonyl chloride, compound 18e was obtained as awhite solid. LC-MS: calcd. for C₁₆H₁₈Cl₂N₂O₂S₂: 404.02 found: 404.7[M+H]⁺.

18f (W₁=5-Cl, R₁=cyclohexyl. R₄/R₄=H, R_(3d)=3,4-dimethoxyphenyl)

To a solution of 15l (0.3 g, 1 mmol) in dry CH₂Cl₂ (5 mL), DIPEA (0.15mL, 1.11 mmol) was added and stirred at 0° C. A solution of3,4-dimethoxy benzenesulfonyl chloride (0.26 g, 1.11 mmol) in dry CH₂Cl₂(3 mL) was added over a period of 8 hrs and stirred for additional for 5hrs at room temperature. The reaction was diluted with CH₂Cl₂ (20 mL)and washed with saturated solution of NaHCO₃ (2×110 mL), water (10 mL)and brine (20 mL). The organic layer was dried over sodium sulfate,filtered and concentrated under vacuum. The crude product was purifiedby flash chromatography on a silica gel column using a mixture ofpetroleum ether and ethyl acetate as eluent to give 18f as semitransparent solid (0.5 g, 90% yield): LC/MS calcd for C₂₃H₃₁ClN₂O₄S₂:498 [M+Na⁺], found: 521.

EXAMPLE 12

General procedure for the preparation of 19: To a solution of Ph₃PMeBr(1.5 equiv.) in dry THF was added t-C₅HIOK (1.5 equiv.) in portionsunder argon. After the mixture was stirred at room temperature for 0.5h, a solution of the corresponding benzophenone derivative 6 in THF wasadded dropwise. The reaction mixture was then stirred at roomtemperature under argon overnight. The reaction mixture was quenchedwith H₂O and was extracted twice with EtOAc. The combined organic layerswere washed with saturated NaHCO₃, brine, dried over Na₂SO₄, filtered,concentrated, and the residue was purified by column chromatography onsilica gel.

Following the general procedure, 19a (W₁=5-Cl, R₁=2-chloroiphenyl) wasobtained as slightly orange oil (1.95 g, 73%) after chromatography(EtOAc/Hexane: 10/90). ¹H NMR (500 MHz, CDCl₃) δ 3.66 (b, 2H), 5.61 (s,1H), 5.87 (s, 1H), 6.57 (d, J=8.4 Hz, 1H), 7.06 (m, 2H), 7.17-7.23 (m,3H), 7.26-7.31 (m, 1H); MS calcd for C₁₄H₁₂Cl₂N (MH⁺) 264.04, found264.0.

Following the general procedure, 19b (W₁=5-Cl, R₁=2-fluorophenyl) wasobtained as colorless oil (5.93 g, 80%) after chromatography(EtOAc/Hexane: 10/90). ¹H NMR (500 MHz, CDCl₃) δ 3.67 (b, 2H), 5.61 (s,1H), 5.86 (s, 1H), 6.62 (d, J=8.3 Hz, 1H), 7.04-7.10 (m, 3H), 7.16-7.19(m, 1H), 7.25-7.35 (m, 2H);

Following the general procedure, 19c (W₁=5-Br, R₁=2-fluorophenyl) wasobtained as colorless oil (7.20 g, 73%) after chromatography(EtOAc/Hexane: 10/90). ¹H NMR (500 MHz, CDCl₃) δ 3.65 (b, 2H), 5.61 (s,1H), 5.87 (s, 1H), 6.57 (d, J=8.3 Hz, 1H), 7.06 (m, 2H), 7.16-7.22 (m,3H), 7.25-7.30 (m, 1H);

Following the general procedure, 19d (W₁=5-Cl, R₁=phenyl) was ascolorless oil (4.26 g, 61%) after chromatography (EtOAc/Hexane: 10/90).¹H NMR (500 MHz, CDCl₃) δ 3.55 (b, 2H), 5.36 (s, 1H), 5.81 (s, 1H), 6.62(d, J=9.1 Hz, 1H), 7.11 (s, 1H), 7.11-7.13 (m, 1H), 7.31-7.37 (m, 4H);General procedure for the preparation of 20: To a solution of thecorresponding styrene derivative 19 and HSCH₂CO₂Me (3.0 equiv.) in1,4-dioxane was added 1,1′azobis(cyclohexanecarbonitrile) (0.1 equiv.).The reaction mixture was then warmed to 80° C. under argon and stirredat that temperature overnight. Additional1,1′-azobis(cyclohexanecarbonitrile) (0.05-0.1 equiv.) was added to thereaction mixture and it was stirred at 80° C. until TLC analysisindicated disappearance of 19. The reaction mixture was diluted withEtOAc and was washed with saturated NaHCO₃, brine, dried over Na₂SO₄,filtered, concentrated, and the residue was purified by columnchromatography on silica gel.

Following the general procedure, 20a (W₁=5-Cl, R₁=2-chlorophenyl) wasobtained as colorless oil (3.15 g, 84%) after chromatography(EtOAc/Hexane: 15/85). ¹H NMR (500 MHz, CDCl₃) δ 3.10-3.30 (m, 2H), 3.18and 3.23 (AB q, J=14.79 Hz, 2H), 3.74 (s, 3H), 4.68 (t, J=7.7 Hz, 1H),6.60 (d, J=8.9 Hz, 1H), 7.02 (t, J=2.0 Hz, 1H), 7.03 (s, 1H), 7.16-7.26(m, 3H), 7.40 (d, J=8.5 Hz, 1H); MS calcd for C₁₇H₁₈Cl₂NO₂S (MH⁺)370.05, found 370.1.

Following the general procedure, 20b (W₁=5-Cl, R₁=2-fluorophenyl) wasobtained as colorless oil (2.05 g, 82%) after chromatography(EtOAc/Hexane: 20/80). ¹H NMR (500 MHz, CDCl₃) δ 3.18 and 3.23 (AB q,J=15.2 Hz, 2H), 3.22-3.34 (m, 2H), 3.74 (s, 3H), 3.77 (b, 2H), 4.54 (t,J=7.8 Hz, 1H), 6.59 (d, J=8.7 Hz, 1H), 7.00-7.26 (m, 6H).

Following the general procedure, 20c (W₁=5-Br, R₁=2-fluorophenyl) wasobtained as pale orange oil (5.11 g, 79%) after chromatography(EtOAc/Hexane: 20/80). ¹H NMR (500 MHz, CDCl₃) δ 3.18 and 3.23 (AB q,J=14.6 Hz, 2H), 3.21-3.34 (m, 2H), 3.74 (s, 3H), 4.53 (t, J=7.8 Hz, 1H),6.55 (d, J=8.3 Hz, 1H), 7.04-7.26 (m, 6H).

Following the general procedure, 20d (W₁=5-Cl, R₁=phenyl) was obtainedas colorless oil (3.60 g, 75%) after chromatography (EtOAc/Hexane:20/80). ¹H NMR (500 MHz, CDCl₃) δ 3.12 and 3.17 (AB q, J=14.5 Hz, 2H),3.21-3.34 (m, 2H), 3.74 (s, 3H), 4.19 (t, J=7.7 Hz, 1H), 6.58 (d, J=8.5Hz, 1H), 7.03 (dd, J=8.3 Hz, 2.7 Hz, 1H), 7.13 (d, J=2.6 Hz, 1H),7.23-7.26 (m, 3H), 7.31-7.34 (m, 2H).

General procedure for the preparation of amide 21: A solution of thecorresponding ester 20 and ammonia (saturated solution in H₂O, 30equiv.) or MeNH₂ (1.0 M solution in MeOH, 10 equiv.) in MeOH was stirredat room temperature until TLC analysis indicated the completedisappearance of 20. The reaction mixture was concentrated under reducedpressure, and the residue was purified by column chromatography onsilica gel.

Following the general procedure, 21a W₁=5-Cl, R₂ 4=2-chlorophenyl,R_(3a)=H, R_(3b)=H) was obtained as white solid (55 mg, 96%) afterchromatography (MeOH/CH₂Cl₂: 2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ 3.16and 3.27 (AB q, J=16.5 Hz, 2H), 3.66 and 3.71 (AB q, J=13.5 Hz, 2H),5.86 (s, 1H), 6.52-6.54 (m, 1H), 6.57 (b, 1H), 7.03-7.04 (m, 2H),7.26-7.29 (m, 1H), 7.32-7.35 (m, 2H), 7.78-7.80 (m, 1H).

Following the general procedure, 21b (W₁=5-Cl, R₁=2-fluorophenl ═H) wasobtained as white solid (90 mg, 96%) after chromatography (MeOH/CH₂Cl₂:2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ 3.14-3.29 (m, 4H), 3.73 (b, 2H),4.51 (t, J=7.8 Hz, 1H), 5.58 (s, 1H), 6.45 (s, 1H), 6.50 (d, J=8.3 Hz,1H), 7.02-7.17 (m, 5H), 7.24-7.27 (m, 1H).

Following the general procedure, 21c (W₁=5-Br, R₁=2-fluorophenyl,R_(3b)=H, R_(3b)=H) was obtained as white solid (117 mg, 93%) afterchromatography (MeOH/CH₂Cl₂: 2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ3.14-3.29 (m, 4H), 3.60-3.80 (b, 2H), 4.49 (t, J=7.8 Hz, 1H), 5.66 (s,1H), 6.46 (s, 1H), 6.55 (d, J=8.4 Hz, 1H), 7.05-7.17 (m, 4H), 7.21-7.26(m, 2H).

Following the general procedure, 21d (W₁=5-Cl, R₁=phenyl, R_(3a)=H,R_(3b)=H) was obtained as white solid (102 mg, 97%) after chromatography(MeOH/CH₂Cl₂: 2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ 3.10-3.31 (m, 4H),4.15 (t, J=7.9 Hz, 1H), 5.60 (s, 1H), 6.43 (s, 1H), 6.58 (d, J=8.3 Hz,1H), 7.04 (dd, J=8.3 Hz, 2.8 Hz, 1H), 7.12 (d, J=2.3 Hz, 1H), 7.22-7.27(m, 3H), 7.31-7.34 (m, 2H).

Following the general procedure, 21e (W₁=5-Cl, R₁=2-chlorophenyl,R_(3a)=CH₃, R_(3b)=H) was obtained as white solid (102 mg, 91%) afterchromatography (MeOH/CH₂Cl₂: 2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ 2.78(d, J=4.9 Hz, 3H), 3.04-3.19 (m, 2H), 3.24 (s, 2H), 3.6-3.8 (b, 2H),4.59 (t, J=7.7 Hz, 1H), 6.58 (m, 1H), 6.60 (d, J=9.2 Hz, 1H), 7.04 (s,1H), 7.05 (m, 1H), 7.11 (dd, J=7.1 Hz, 2.7 Hz, 1H), 7.20-7.26 (m, 2H),7.40 (dd, J=7.0 Hz, 1.5 Hz, 1H).

Following the general procedure, 21f (W₁=5-Cl, R₁=2-fluorophenyl.R_(3a)=CH₃, R_(3b)=H) was obtained as white solid (91 mg, 96%) afterchromatography (MeOH/CH₂Cl₂: 2/98). ¹H NMR (500 MHz, CDCl₃) δ 2.80 (d,J=5.0 Hz, 3H), 3.09-3.25 (m, 4H), 3.75 (s, 2H), 4.46 (t, J=7.8 Hz, 1H),6.56 (m, 1H), 6.60 (d, J=8.3 Hz, 1H), 7.02-7.14 (m, 5H), 7.25-7.26 (m,1H).

Following the general procedure, 21g (W₁=5-Br, R₁=2-fluorophenyl,R_(3a=CH) ₃, R_(3b)=H) was obtained as white solid (116 mg, 96%) afterchromatography (MeOH/CH₂Cl₂: 2/98). ¹H NMR (500 MHz, CDCl₃) δ 2.80 (d,J=5.0 Hz, 3H), 3.09-3.25 (m, 4H), 3.76 (b, 2H), 4.45 (t, J=7.8 Hz, 1H),6.55 (d, J=8.5 Hz, 1H), 7.05-7.26 (m, 6H).

Following the general procedure, 21h (W₁=5-Cl, R₁=phenyl, R_(3a)=CH₃,R_(3b)=H) was obtained as white solid (86 mg, 96%) after chromatography(MeOH/CH₂Cl₂: 2/98). ¹H NMR (500 MHz, CDCl₃) δ 2.78 (d, J=5.1 Hz, 3H),3.06-3.28 (m, 4H), 4.11 (t, J=7.9 Hz, 1H), 6.51 (m, 1H), 6.58 (d, J=8.3Hz, 1H), 7.04 (dd, J=8.4 Hz, 2.9 Hz, 1H), 7.11 (d, J=2.1 Hz, 1H),7.21-7.22 (m, 2H), 7.25-7.28 (m, 1H), 7.31-7.34 (m, 2H).

Following the general procedure, 21i (W₁=5-Br, R₁=2-fluorophenyl,R_(3a)/R_(3b)=—(CH₂)₂O(CH₂)₂—) was obtained as white solid (55 mg, 95%).MS calcd for C₂₀H₂₃BrFN₂O₂S (MH⁺) 453.07, found 453.0.

Preparation of 21i (W₁=5-Br, R₁=2-fluorophenyl, R₁ =OH, R _(3b)=H. To asolution of the corresponding ester 20c (228 mg, 0.57 mmol), NH₂OHHCl(0.80 g, 11.5 mmol) in dry MeOH was added MeONa (25% wt. in MeOH, 4.0ml). NaCl precipitated from the solution immediately. The reactionmixture was then refluxed overnight. After it was cooled down to roomtemperature, the solution was neutralized with 1.0 N HCl to pH around8.0, and extracted twice with CH₂Cl₂. The combined organic layers werewashed with saturated NaHCO₃, brine, dried over Na₂SO₄, filtered,concentrated, and the residue was purified by column chromatography onsilica gel (MeOH/CH₂Cl₂: 10/90) to give 211 (82 mg, 36%) as pale yellowsolid. ¹H NMR (500 MHz, CDCl₃) δ 3.11-3.29 (m, 4H), 4.50 (m, 1H), 6.60(d, J=7.5 Hz, 1H), 7.06 (t, J=8.9 Hz, 1H), 7.08-7.26 (m, 5H); MS calcdfor C₁₆H₂₇BrFN₂O₂S (MH⁺) 399.02, found 398.9

General procedure for the preparation of 22: To a solution of thecorresponding ester 20 and acetimidoxime hydrochloride salt (5.0 equiv.)in dry THF/MeOH (1:1) was added dropwise a solution of MeONa (25% wt.solution in MeOH, 12.0 equiv.) under argon at room temperature. NaClprecipitated immediately from the reaction mixture. The suspension wasthen stirred at room temperature overnight. After TLC analysis indicatedthe completion of the reaction, the reaction mixture was quenched withH₂O and was extracted twice with EtOAc. The combined organic layers werewashed with brine, dried over Na₂SO₄, filtered, concentrated, and theresidue was purified by column chromatography on silica gel.

Following the general procedure, 22a (W₁=5-Cl, R₁=2-chlorophenyl) wasobtained as colorless oil (175 mg, 56%) after chromatography(EtOAc/Hexane: 20/80). ¹H NMR (500 MHz, CDCl₃) δ 2.41 (s, 3H), 3.19-3.33(m, 2H), 3.65-3.85 (b, 2H), 3.79 and 3.83 (AB q, J=15.4 Hz, 2H), 4.69(t, J=7.7 Hz, 1H), 6.60 (d, J=8.3 Hz, 1H), 7.01-7.04 (m, 2H), 7.15-7.16(m, 1H), 7.21-7.26 (m, 2H), 7.39 (d, J=8.6 Hz, 1H).

Following the general procedure 22b (W₁=5-Cl, R₁=2-fluorophenyl) ascolorless oil (101 mg, 42%) after chromatography (EtOAc/Hexane: 20/80).¹H NMR (500 MHz, CDCl₃) δ 2.41 (s, 3H), 3.23-3.37 (m, 2H), 3.76 and 3.82(AB q, J=15.5 Hz, 2H), 4.54 (t, 7.8 Hz, 1H), 6.60 (d, J=8.5 Hz, 1H),7.01-7.27 (m, 6H).

Following the general procedure, 22c (W₁=5-Br, R₁=2-fluorophenyl) ascolorless oil (137 mg, 45%) after chromatography (EtOAc/Hexane: 20/80).¹H NMR (500 MHz, CDCl₃) δ 2.41 (s, 3H), 3.23-3.37 (m, 2H), 3.76 and 3.81(AB q, J=15.4 Hz, 2H), 4.53 (t, 7.8 Hz, 1H), 6.55 (d, J=8.4 Hz, 1H),7.04-7.27 (m, 6H).

Following the general procedure, 22d (W₁=5-Cl, R₁=flurophenyl) ascolorless oil (146 mg, 51%) after chromatography (EtOAc/Hexane: 30/70).¹H NMR (500 MHz, CDCl₃) δ 2.41 (s, 3H), 3.23-3.37 (m, 2H), 3.66 and 3.73(AB q, J=14.9 Hz, 2H), 4.16 (t, 7.7 Hz, 1H), 6.58 (d, J=8.3 Hz, 1H),7.04 (dd, J=8.3 Hz, 2.5 Hz, 1H), 7.14 (d, J=2.5 Hz, 1H), 7.22-7.27 (m,3H), 7.31-7.34 (m, 2H).

EXAMPLE 13

Preparation of 25a (W₁=H, R₁=3-chlorophenyl): To a stirred suspension ofNaH (0.60 g, 60% purity, 15.0 mmol) in dry DMF was added dropwise methyl3-10 chlorophenylacetate (24a, R₁=3-chlorophenyl) (2.22 g, 12.0 mmol)via syringe at 0° C. under argon. After the solution was stirred at 0°C. for 0.5 hrs, 1,2-dinitrobenzene (3a, R₁=H)(1.68 g, 10.0 mmol) wasadded in small portions. The reaction mixture was then stirred at 0° C.and allowed to warm to rt overnight. The reaction was quenched withsaturated aqueous NH₄Cl solution and was extracted twice with EtOAc. Thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered, concentrated, and the residue was purified by columnchromatography on silica gel (EtOAc/Hexane: 10/90) to give 25a, whichwas further purified by recrystallization from EtOAc/Hexane, as a yellowsolid (1.28 g, 42%). ¹H NMR (500 MHz, CDCl₃) δ 3.79 (s, 3H), 5.08 (s,1H), 7.19-7.21 (m, 1H), 7.30-7.31 (m, 3H), 7.52 (t, J=7.9 Hz, 1H), 7.66(d, J=7.5 Hz, 1H), 8.16 (d, J=8.4 Hz, 1H), 8.18 (s, 1H); MS calcd forC₁₅H₁₃ClNO₄ (MH⁺) 306.06, found 306.0.

25b (W₁=H, R₁=2-methylphenyl)

Using a similar procedure as for 25a, 25b was obtained from ethyl1-methylphenylacetate (6.72 g, 40.0 mmol) and 1,2-dinitrobenzene (10.70g, 60.0 mmol) as a yellow solid (2.63 g, 22%). ¹H NMR (500 MHz, CDCl₃) δ1.27 (t, J=7.1 Hz, 3H), 2.28 (s, 3H), 4.26 (q, J=7.1 Hz, 2H), 5.30 (s,1H), 7.23-7.28 (m, 5H), 7.60 (dd, J=8.2 Hz, 1.7 Hz, 1H), 7.78 (d, J=1.7Hz, 1H), 7.87 (d, J=8.3 Hz, 1H).

25c=(W₁=H, R₁ =methylphenyl)

Using a similar procedure as for 25a, 25c was obtained from methylphenylacetate (2.68 g, 17.8 mmol), and 1,2-dinitrobenzene (2.50 g, 14.9mmol) as yellow solid (1.26 g, 31%) after chromatography on silica gel(EtOAc/Hexane: 10/90). ¹H NMR (500 MHz, CDCl₃) δ 3.70 (s, 3H), 5.04 (s,1H), 7.18-7.32 (m, 3H), 7.40-7.49 (m, 2H), 7.58 (t, J=7.6 Hz, 1H), 7.72(d, J=7.8 Hz, 1H), 8.07 (t, J=7.8 Hz, 1H).

25d (W₁=H, R₁=1-naphthyl)

Using a similar procedure as for 25a, 25d was obtained from methyl1-naphthalenylacetate (6.01 g, 30.0 mmol) and 1,2-dinitrobenzene (3.36g, 20.0 mmol) as a yellow solid (1.63 g, 25%). ¹H NMR (500 MHz, CDCl₃) δ3.81 (s, 3H), 5.89 (s, 1H), 7.42-7.45 (m, 1H), 7.51-7.55 (m, 3H), 7.66(dd, J=8.3 Hz, 1.9 Hz, 1H), 7.80-7.93 (m, 6H).

General procedure for the preparation of 26: A solution of 25 andammonia (saturated solution in H₂O, 30 equiv.) or MeNH₂ (1.0 M solutionin MeOH, 10 equiv.) in MeOH, or PhCH₂CH₂NH₂ (neat, 10 equiv.) wasstirred at indicated temperature until TLC analysis indicated thecomplete disappearance of 25. The reaction mixture was concentratedunder reduced pressure, and the residue was purified by columnchromatography on silica gel

Following the general procedure, 26a (W₁=H, R₁=3-chlorophenyl, R=CH₃)was obtained from 25a (120 mg, 0.39 mmol), and MeNH₂ (1.0 M solution inMeOH, 3.9 ml, 3.9 mmol) as pale yellow solid (112 mg, 94%) afterchromatography (EtOAc/Hexane: 40/60). ¹H NMR (500 MHz, CDCl₃) δ 2.88 (d,J=4.9 Hz, 3H), 4.90 (s, 1H), 5.72 (b, 1H), 7.18-7.20 (m, 1H), 7.26-7.31(m, 3H), 7.52 (t, J=8.0 Hz, 1H), 7.66 (d, J=8.1 Hz, 1H), 8.14 (s, 1H),8.15 (d, J=7.3 Hz, 1H); MS calcd for C₁₅H₁₄ClN₂O₃ (MH⁺) 305.07, found305.1.

Following the general procedure, 26b (W₁=H, R₁=3-chlorophenyl,R=2-phenylethyl) was obtained from 25a (270 mg, 0.88 mmol), andPhCH₂CH₂NH₂ (1.07 g, 8.8 mmol) as pale yellow oil (310 mg, 89%) afterchromatography (EtOAc/Hexane: 30/70). ¹H NMR (500 MHz, CDCl₃) δ 2.81 (t,J=6.7 Hz, 2H), 3.54-3.64 (m, 2H), 4.82 (s, 1H), 5.62 (m, 1H), 7.05-7.09(m, 3H), 7.20-7.30 (m, 6H), 7.48 (t, J=7.8 Hz, 1H), 7.55 (d, J=7.5 Hz,1H), 8.08 (s, 1H), 8.13 (d, J=8.2 Hz, 1H); MS calcd for C₂₂H₂₀ClN₂O₃(MH⁺) 395.12, found 395.1.

Following the general procedure, 26c (W₁=H, R₁=3-chlorophenyl, R=H) wasobtained from 25a (225 mg, 0.74 mmol), and NH₄OH (−14.8 N in H₂₀, 1.5ml, 22.2 mmol) as pale yellow solid (194 mg, 91%) after chromatography(MeOH/CH₂Cl₂: 2.5/97.5). ¹H NMR (500 MHz, CDCl₃) δ 4.97 (s, 1H), 5.65(s, 1H), 5.86 (s, 1H), 7.20-7.22 (m, 1H), 7.30-7.32 (m, 3H), 7.53 (t,J=8.2 Hz, 1H), 7.66 (d, J=7.7 Hz, 1H), 8.15-8.16 (m, 1H), 8.17 (s, 1H);

General procedure for the preparation of 27: To a solution of thecorresponding amide derivative 26 in dry THF was added dropwise asolution of BH₃.THF complex in THF (1.0 M, 2-3 equiv.) at 0° C. underargon. The reaction mixture was then stirred at 0° C. and allowed towarm to rt overnight. After the reaction was complete indicated by TLCanalysis, the reaction mixture was carefully quenched with MeOH at 0°C., and then diluted with EtOAc. The organic layer was washed withsaturated NaHCO₃, brine, dried over Na₂SO₄, filtered, concentrated, andthe residue was purified by column chromatography on silica gel.

Following the general procedure, 27a (W₁=H, R₁=3-chlorophenyl, R=CH₃)was obtained from 26a (129 mg, 0.42 mmol), and BH₃-THF complex in THF(1.0 M, 1.26 ml, 1.26 mmol) as pale yellow solid (110 mg, 89%) afterchromatography (EtOAc/Hexane: 20/80). ¹H NMR (500 MHz, CDCl₃) δ 2.64(dd, J=6.0 Hz, 2.0 Hz, 3H), 3.24-3.51 (m, 2H), 4.71 (t, J=7.4 Hz, 1H),4.83-4.86 (m, 1H), 7.15-7.37 (m, 3H), 7.53-7.65 (m, 3H), 8.07-8.16 (m,2H); MS calcd for C₁₅H₁₆ClN₂O₂ (MH⁺) 291.09, found 291.1.

Following the general procedure, 27b (W₁=H, R₁=3-chlorophenyl,R=2-phenylethyl) was obtained from 26b (196 mg, 0.50 mmol), and BH₃-THFcomplex in THF (1.0 M, 1.99 ml, 1.99 mmol) as pale yellow oil (147 mg,88%) after chromatography (EtOAc/Hexane: 10/90). MS calcd forC₂₂H₂₂ClN₂O₂ (MH⁺) 381.04, found 381.2.

General procedure for the preparation of 28: To a solution of thecorresponding amine derivative 27 and DIEA (2.0-5.0 equiv.) in dry THFwas added dropwise BrCH₂CO₂Me (1.2-2.5 equiv.) at 0° C. under argon. Thereaction mixture was then stirred at 0° C. and allowed to warm to rtovernight. The reaction mixture was quenched with saturated NaHCO₃ andwas extracted twice with EtOAc. The combined organic layers were washedwith saturated NaHCO₃, brine, dried over Na₂SO₄, filtered, concentrated,and the residue was purified by column chromatography on silica gel.

Following the general procedure, 28a (W₁=H, R₁=3-chlorophenyl, R=CH₃)was obtained from 27a (110 mg, 0.38 mmol), BrCH₂CO₂Me (116 mg, 0.76mmol), and DIEA (245 mg, 1.90 mmol) as colorless oil (104 mg, 76%) afterchromatography (EtOAc/Hexane: 20/80). ¹H NMR (500 MHz, CDCl₃) δ 2.43 (s,3H), 3.17-3.22 (m, 2H), 3.28 and 3.32 (AB q, J=15.5 Hz, 2H), 3.70 (s,3H), 4.26 (t, J=7.2 Hz, 1H), 7.12 (d, J=6.6 Hz, 1H), 7.21-7.26 (m, 3H),7.48 (t, J=7.7 Hz, 1H), 7.60 (d, J=7.7 Hz, 1H), 8.09 (d, J=7.7 Hz, 1H),8.14 (s, 1H); MS calcd for C₁₈H₂₀ClN₂O₄ (MH⁺) 363.11, found 363.1.

Following the general procedure, 28b (W₁=H, R₁=3-chlorophenyl,R=2-phenylethyl) was obtained from 27b (70 mg, 0.18 mmol), BrCH₂CO₂Me(56 mg, 0.37 mmol), and DIEA (119 mg, 0.92 mmol) as colorless oil (81mg, 97%) after chromatography (EtOAc/Hexane: 10/90). ¹H NMR (500 MHz,CDCl₃) δ 2.61-2.65 (m, 2H), 2.94 (t, J=7.5 Hz, 2H), 3.31-3.33 (m, 2H),3.36 (s, 2H), 3.69 (s, 3H), 4.16 (t, J=7.8 Hz, 1H), 7.04-7.07 (m, 3H),7.15-7.26 (m, 6H), 7.43 (t, J=8.1 Hz, 1H), 7.48 (d, J=7.3 Hz, 1H),8.06-8.07 (m, 2H); MS calcd for C₂₅H₂₆ClN₂O₄ (MH⁺) 453.16, found 453.3.

Following the general procedure, 28c (W₁=H, R₁=3-chlorophenyl, R=H) wasobtained from 27c (35 mg, 0.13 mmol), BrCH₂CO₂Me (77 mg, 0.50 mmol), andDIEA (163 mg, 1.26 mmol) as colorless oil (50 mg, 94%) afterchromatography (EtOAc/Hexane: 30/70). MS calcd for C₂₀H₂₂ClN₂O₆ (MH⁺)421.12, found 421.1.

General procedure for the preparation of 30: A suspension of thecorresponding 4 and Pd/C (10% on the charcoal, 0.05 equiv.) in EtOAc wasstirred under an atmospheric of hydrogen at atmospheric pressure for 2h. The Pd/C was filtered off. The filtration was concentrated, and theresidue was purified by column chromatography on silica gel.

Following the general procedure, 30a (W₁=H, R₁=3-chlorophenyl, R=CH₃)was obtained from 28a (15 mg, 0.04 mmol) and Pd/C as colorless oil (13mg, 94%) after chromatography (EtOAc/Hexane: 50/50). ¹H NMR (500 MHz,CDCl₃) δ 2.42 (s, 3H), 3.09-3.16 (m, 2H), 3.27 and 3.31 (AB q, J=15.5Hz, 2H), 3.61 (b, 2H), 3.69 (s, 3H), 4.04 (t, J=7.7 Hz, 1H), 6.52-6.53(m, 2H), 6.63 (d, J=8.1 Hz, 1H), 7.07 (t, J=7.9 Hz, 1H), 7.15-7.26 (m,4H); MS calcd for C₁₈H₂₂ClN₂O₂ (MH⁺) 333.14, found 333.1.

Following the general procedure, 30b (W₁=H, R₁=3-chlorophenyl,R=2-phenylethyl) was obtained from 28b (32 mg, 0.07 mmol) and Pd/C ascolorless oil (27 mg, 90%) after chromatography (EtOAc/Hexane: 30/70).¹H NMR (500 MHz, CDCl₃) δ 2.63-2.67 (m, 2H), 2.91 (t, J=7.6 Hz, 2H),3.24-3.32 (m, 2H), 3.34 (s, 2H), 3.59 (b, 2H), 3.66 (s, 3H), 3.98 (t,J=7.6 Hz, 1H), 6.49 (d, J=1.7 Hz, 1H), 6.52 (dd, J=8.0 Hz, 1.8 Hz, 1H),6.60 (d, J=7.4 Hz, 1H), 7.05-7.14 (m, 4H), 7.16-7.20 (m, 3H), 7.22-7.26(m, 3H); MS calcd for C₂₅H₂₈ClN₂O₂ (MH⁺) 423.19, found 423.1.

General procedure for the preparation of 29 and 31: A solution of thecorresponding ester derivative 28 (or 30) in cyclohexylamine was stirredat 120° C. overnight. Cyclohexylamine was then removed under reducedpressure, and the residue was purified by column chromatography onsilica gel.

Following the general procedure, 29a (W₁=H, R₁=3-chlorophenyl, R=CH₃,R₄=cyclohexyl) was obtained from 28a (11 mg, 0.03 mmol) as pale brownoil (12.9 mg, 99%) after chromatography (EtOAc/Hexane: 50/50). ¹H NMR(500 MHz, CDCl₃) δ 0.77-0.82 (m, 2H), 1.07 (m, 1H), 1.24-1.31 (m, 3H),1.53-1.55 (m, 2H), 1.64-1.67 (m, 2H), 2.33 (s, 3H), 3.07 (s, 2H), 3.11(d, J=8.1 Hz, 2H), 3.59-3.61 (m, 1H), 4.22 (t, J=8.1 Hz, 1H), 6.37 (bd,J=8.0 Hz, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.20 (s, 1H), 7.24-7.31 (m, 2H),7.49-7.55 (m, 2H), 8.10-8.13 (m, 2H); MS calcd for C₂₃H₂₉ClN₃O₃ (MH⁺)430.19, found 430.1.

29b (W₁=H, R₁=3-chlorophenyl, R=2-phenylethyl, R=cyclohexyl): Followingthe general procedure, 28b (4.0 mg, 0.0088 mmol) was employed to give29b (4.4 mg, 95%) as pale yellow oil after chromatography (EtOAc/Hexane:40/60). MS calcd for C₃₀H₃₅ClN₃O₃ (MH⁺) 520.24, found 520.2.

31a (W₁=H, R₁=3-chlorophenyl, R=CH₃, R_(3a)=cyclohexyl): Following thegeneral procedure, 30a (5.0 mg, 0.015 mmol) was employed to give 31a(5.5 mg, 92%) as pale yellow oil after chromatography (EtOAc/Hexane:60/40). LC-MS calcd for C₂₃H₃₁ClN₃O (MH⁺) 400.22, found 400.2.

EXAMPLE 14 Further Synthesis of Representative Compounds

34a (W₁=5-Cl, R₁=2-chlorophenyl, R_(2b)/R_(2c)=CH₃)

To a stirred solution of 11s (100 mg, 0.280 mmol) in THF (2.0 ml) undera nitrogen atmosphere at rt was added acetic acid (20 μL, to give a 1%solution in THF) and paraformaldehyde (100 mg) and the reaction mixturewas heated to 60° C. for 24 hrs. Further acetic acid (20 μl) andparaformaldehyde (100 mg) were added and stirring was continued for 48hrs at 60° C. Sodium cyanoborohydride (35.2 mg, 0.56 mmol, 2eq) wasadded and stirring was continued at 60° C. for a further 24 hrs. Thereaction mixture was partitioned between dichloromethane and brine, theaqueous phase was back extracted, then the combined organic phase wasdried with brine and sodium sulfate, then filtered and evaporated togive the crude N,N-dimethylamine (130 mg) as a yellow oil. The crudematerial was purified by flash chromatography on silica (10 g) withpetroleum ether:ethyl acetate (10:1 then 5:1) to give 34a as a yellowoil (53.0 mg, 0.138 mmol, 64% yield). R_(f) (silica, petroleumether:ethyl acetate (5:1))=0.72. HPLC (214 nm) t_(R)=10.13 (92.0%) min.¹H NMR (400 MHz, CDCl₃) δ 2.58 (s, 6H), 3.17 (d, J=1.2 Hz, 2H), 3.64 (s,3H), 6.42 (s, 1H), 7.12-7.28 (m, 4H), 7.37 (dd, J=8.0, 1.2 Hz, 1H), 7.57(d, J=2.8 Hz, 1H), 7.60 (dd, J=8.4, 1.6 Hz, 1H). ¹³C NMR (400 MHz,CDCl₃) 8.34.5, 45.1, 45.2, 52.1, 123.0, 126.9, 128.4, 128.5, 129.5,129.6, 129.9, 134.6, 137.0, 137.8, 152.0, 170.0. ESMS m/z 384.3[(M+H)]⁺. LC/MS t_(R) 9.31 (383.9 [M+H]⁺) min.

34b (W₁=5-Cl, R₁=2-chlorophenyl, R_(2b)=CH₃CO, R_(2c)=H)

To a stirred solution of 11s (50 mg, 0.140 mmol) in THF (5.0 ml) under anitrogen atmosphere at rt was added acetic anhydride (26.5 μL, 0.280mmol, 2eq), DIEA (90.6 μL, 0.702 mmol, 5eq) and DMAP (1.7 mg, 0.014mmol, 0.1 eq) and stirring was continued for 1 hr. Acetyl chloride (20μL, 0.28 mmol, 2eq) was added and was stirring continued for 1 hr.Further acetyl chloride (20 μL, 0.28 mmol, 2eq) was added and stirringwas continued for 24 hrs. The reaction mixture was partitioned betweendichloromethane and water, the aqueous phase was back extracted twicewith dichloromethane, then the combined organic phases were dried withbrine and sodium sulfate, then filtered and evaporated to give the crudeamide (120 mg) as a yellow oil. The crude material was purified by flashchromatography on silica (5 g) with petroleum ether:ethyl acetate (2:1)to give 34b as a white solid (34.0 mg, 0.853 mmol, 61% yield). R_(f)(silica, petroleum ether:ethyl acetate (2:1))=0.20. HPLC (214 nm)t_(R)=8.82 (87.7%) min. ¹H NMR (400 MHz, CDCl₃) δ 2.38 (s, 3H), 3.15 (d,J=17.2 Hz, 1H), 3.22 (d, J=17.2 Hz, 1H), 3.78 (s, 3H), 5.90 (s, 1H),6.78 (brs, 1H), 7.24 (dd, J=8.8, 2.4 Hz, 1H), 7.29-7.37 (m, 1H),7.39-7.47 (m, 2H), 8.01 (d, J=7.6 Hz, 1H), 8.07 (d, J=8.4 Hz, 1H), 5.38(brs, 1H). ³C NMR (400 MHz, CDCl₃) 8.24.3, 33.4, 45.5, 52.9, 125.1,127.4, 127.7, 128.4, 129.5, 130.2, 130.3, 130.6, 134.8, 134.9, 169.4,171.6. ESMS m/z 292.3 [(M−HSCH₂CO₂CH₃+H)]⁺, 398.1 [(M+H)]⁺. LC/MS t_(R)8.73 (397.9 [(M+H)]⁺, 794.8 [(2M+H)]⁺) min.

34c (W₁=H, R₁=2-methylphenyl, R_(2b)=benzoyl, R_(2c)=H)

Step 1. To a solution ofbenzophenone 2 (W₁=H, R₁=2-methylphenyl) (287mg, 0.9 mmol) in DMF (10 ml) at 0° C. was added sodium borohydride (35mg, 0.9 mmol) in one portion and the mixture stirred for 2 h, keepingthe temperature between 0-4° C. The mixture was then poured into water(200 ml), acidified to pH=5 with dilute HCl, then extracted with ethylacetate (20 ml). The aqueous layer was extracted with ethyl acetate (20ml×3) and the combined ethyl acetate layers were washed with water (60ml×2). The organic phase was dried (Na₂SO₄) and concentrated underreduced pressure to yield the crude product that is used immediately inthe next step.

Step 2: To a stirred solution of the above crude product in TFA (5.0 ml)under a nitrogen atmosphere at rt was added methyl thioglycolate (0.323ml, 3.62 mmol, 4eq). After stirring for 18 hrs the TFA was evaporatedand the residue partitioned between dichloromethane and aqueous NaOH (1mol/L). The aqueous phase was back extracted with dichloromethane andthe combined organics were dried with brine and sodium sulphate, thenfiltered and evaporated to give the crude product (317 mg) as a yellowsolid: The crude material was purified by flash chromatography on silica(15 g) by eluting with petroleum ether:ethyl acetate (5:1 then 2:1) togive the thiol ether 34c as a yellow oil (225 mg, 0.555 mmol, 61%):R_(f) (petroleum ether:ethyl acetate (2:1)=0.65; EM_(calc.)=405.1,(M+1)⁺ _(obs)=406.1; ¹HNMR (400 MHz) 8.90 (1H, br s), 8.01 (1H, d, J=7.6Hz), 7.88-7.96 (2H, m), 7.70 (1H, dd, J=0.8, 7.6 Hz), 7.40-759 (m, 3H),7.01-7.36 (6H, m), 5.83 (1H, s), 3.41 (3H, s), 3.13 (2H, s), 2.11 (3H,s); ¹³CNMR (400 MHz) 170.79, 165.98, 137.35, 136.22, 135.46, 134.76,133.50, 131.77, 131.11, 130.12, 129.08, 128.55, 128.49, 128.44, 127.89,127.53, 126.46, 125.26, 124.61, 52.25, 46.41, 33.27, 19.12

34d (W₁=5-Cl, R₁=3-methyl-2-thiophenyl, R_(2b)=—NHC(═O)C(CH₃)₃,R_(2c)=H)

To a solution of the alcohol 3c (W₁=5-Cl, R₁=3-methyl-2-thiophenyl) (50mg, 0.14 mmol) in methanol (3 ml) was added 1M HCl (3 ml) dropwise. Thereaction was then heated at a gentle reflux for 15 min. After this timeTLC analysis indicated the formation of a new product. The reaction wasworked up by diluting with brine (30 ml), raising the pH to >10 with 1MNaOH and the aqueous solution was extracted with dichloromethane (4×20ml). The combined organic phase was dried over sodium sulfate, filteredand the solvent removed in vacuo. The residue was dissolved indichloromethane (5 ml), methyl thioglycolate (50 μL) was added followedby TFA (50 μL). After 15 min TLC indicated the complete consumption ofstarting material. The solvent was removed in vacuo and the residue waspurified on silica gel (50 g) using petroleum spirit/ethyl acetate 4:1as eluent to yield 34d, isolated as white solid (50 mg, 79% yield);clogP=5.65; Rfpetroleum spirit/ethyl acetate, 4:1)=0.50; HPLC (214 nm)t_(R)=10.81 (99.34%) min; ¹H NMR (400 MHz, CDCl₃) δ 1.33 (s, 9H), 2.10(s, 3H), 3.15 (d, J=15.6 Hz, 1H), 3.27 (d, J=15.6 Hz, 1H), 3.71 (s, 3H),5.83 (s, 1H), 6.83 (d, J=5.1 Hz, 1H), 7.20 (d, J=5.1 Hz, 1H), 7.25 (dd,J=2.4, 8.8 Hz, 1H), 7.43 (d, J=2.4 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 8.22(s, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 13.9, 27.5, 33.4, 39.7, 42.6, 52.5,124.6, 126.6, 128.5, 128.9, 130.6, 130.8, 132.4, 134.5, 135.0, 136.2,170.8, 177.3; ESMS mnz 426.3 [M+H]⁺, 443.3 [M+NH₄]⁺; LC/MS t_(R)=9.74(426.2 [M+H]⁺, 851.2 [2M+3H]⁺) min.

34e (W₁=5-Cl, R₁=1-methyl-2-pyrrolyl, R_(2b)=—NHC(═O)C(CH₃)₃, R_(2c)=H)

Following a similar procedure of 34c, 34e was obtained from the reactionof methyl thioglycolate and the condensation product of 2a with1-methylpyrrole-2-carboxaldehyde. Compound 34e was isolated as a browncrystaline solid (233 mg, 75% yield); clogP=3.90; R_(f) (petroleumether:ethyl acetate (4:1)=0.53; HPLC (214 nm) t_(R) _(=9.51) (90.86%)min; ¹H NMR (400 MHz, CDCl₃) δ 1.27 (s, 9H), 3.10 (d, J=15.5 Hz, 1H),3.27 (d, J=15.5 Hz, 1H), 3.50 (s, 3H), 3.70 (s, 3H), 5.54 (s, 1H), 6.07(m, 1H), 6.14 (m, 1H), 6.62 (t, J=2.2 Hz, 1H), 7.26 (m, 1H), 7.33 (d,J=2.4 Hz, 1H), ), 7.82 (d, J=8.6 Hz, 1H), 8.16 (s, 1H); ¹³C NMR (100MHz, CDCl₃) δ 27.4, 32.8, 34.0, 39.6, 42.2, 52.5, 107.3, 110.0, 124.0,126.5, 127.3, 128.5, 129.4, 130.4, 131.5, 134.8, 170.9, 177.2; ESMS m/z303.2 [M−SCH₂CO₂CH₃]⁺, 408.9 [M+H]⁺; LC/MS t_(R)=9.37 (303.1[M−SCH₂CO₂CH₃]⁺, 408.9 [M+H]⁺, 817.1 [2M+H]⁺) min.

Preparation of 35f (W₁=5-Cl, R₁=2-fluorophenyl, R_(3a)=CH₃)

To a solution of the corresponding amide derivative 21f (178 mg, 0.48mmol) in dry THF was added dropwise a solution of BH₃.THF complex in THF(1.0 M, 1.45 ml, 1.45 mmol) at 0° C. under argon. The reaction mixturewas then stirred at 0° C.-rt overnight. After the reaction was completeindicated by TLC analysis, the reaction mixture was carefully quenchedwith MeOH at 0° C., and then diluted with EtOAc. The organic layer waswashed with saturated NaHCO₃, brine, dried over Na₂SO₄, filtered,concentrated, and the residue was purified by column chromatography onsilica gel (EtOAc/Hexane: 30170) to give 35f (138 mg, 84%) as colorlessoil. ¹H NMR (500 MHz, CDCl₃) δ 2.50-2.52 (m, 3H), 2.67-2.80 (m, 2H),2.94-3.00 (m, 2H), 3.11-3.26 (m, 2H), 3.63-3.68 (b, 2H), 4.05-4.08 (m,1H), 4.42-4.47 (m, 1H), 6.60-6.63 (m, 1H), 7.03-7.27 (m, 6H); MS calcdfor C₁₇H₂₁ClFN₂S (MH⁺) 339.11, found 339.1.

Preparation of 36a (W₁=5-Br, R₁=2-fluorophenyl, X_(a)=S): A solution ofthe corresponding ester 20c (175 mg, 0.44 mmol) and KSCN (215 mg, 2.21mmol) in AcOH/H₂O (3:1) was heated at 80° C. overnight. After it wascooled down to room temperature, the reaction mixture was then dilutedwith H₂O, and was extracted twice with EtOAc. The combined organiclayers were washed with saturated NaHCO₃, brine, dried over Na₂SO₄,filtered, concentrated, and the residue was purified by columnchromatography on silica gel (MeOH/CH₂Cl₂, 2.5/97.5) to give 36a (88 mg,44%) as white solid. MS calcd for C₁₈H₁₉BrFN₂O₂S₂ (MH⁺) 457.01, found456.9.

Preparation of 36b (W₁=5-Br, R₁=2-fluorophenyl, X_(a)=O): A solution ofthe corresponding ester 20c (131 mg, 0.33 mmol) and KOCN (135 mg, 1.66mmol) in AcOH/H₂O (3:1) was heated at 80° C. overnight. After it wascooled down to room temperature, the reaction mixture was then dilutedwith H₂O, and was extracted twice with EtOAc. The combined organiclayers were washed with saturated NaHCO₃, brine, dried over Na₂SO₄,filtered, concentrated, and the residue was purified by columnchromatography on silica gel (MeOHVCH₂Cl₂, 2.5/97.5) to give 36b (90 mg,62%) as white solid. ¹H NMR (500 MHz, CDCl₃) δ 3.23 (d, J=7.9 Hz, 2H),3.28 (s, 2H), 3.82 (s, 3H), 4.75 (t, J=7.9 Hz, 1H), 7.02 (t, J=9.4 Hz,1H), 7.16-7.20 (m, 2H), 7.25-7.29 (m, 2H), 7.33 (dd, J=8.4 Hz, 1.6 Hz,1H), 7.60 (d, J=8.4 Hz, 1H), 8.66 (b, 1H); MS calcd for C₁₈HIgBrFN₂O₃S(MH⁺) 441.03, found 441.0.

Preparation of 36c (W₁=5-Br, R₁=2-fluorophenyl, X=NH): A solution of thecorresponding ester 20c (186 mg, 0.47 mmol), HCl (1.0 M in Et₂₀, 1.0 ml,1.0 mmol), and H₂NCN (80 mg, 2.0 mmol) in chlorobenzene was stirred at130° C. overnight. The reaction mixture was then concentrated underreduced pressure, and the product was precipitated from the solution.The white solid was collected by filtration, and washed with ether togive 36c (115 mg, 52%). ¹H NMR (500 MHz, CDCl₃) δ 1.88 (b, 3H),3.24-3.35 (m, 4H), 3.67 (s, 3H), 4.78 (t, J=7.9 Hz, 1H), 6.99 (t, J=9.3Hz, 1H), 7.10 (d, J=8.2 Hz, 1H), 7.15 (t, J=7.7 Hz, 1H), 7.22-7.26 (m,1H), 7.33 (t, J=7.7 Hz, 1H), 7.41-7.42 (m, 1H), 7.43 (s, 1H); MS calcdfor C₁₈H₂₀BrFN₃O₂S (MH⁺) 440.04, found 440.1.

Preparation of 37a (R₁=5-Br, R₂₌₂-fluorophenyl): To a solution of1,2-phenylenediamine (70 mg, 0.65 mmol) in dry THF was added dropwise asolution of n-BuLi (2.0 M, 1.0 ml, 2.0 mmol) in cyclohexane at 0° C.under argon. The solution was stirred at 0° C. for 0.5 hrs. To thismixture was then added dropwise a solution of 20c (130 mg, 0.33 mmol) inTHF. The reaction mixture was stirred at 0° C.-rt overnight. After thereaction was complete indicated by LC-MS analysis, the reaction mixturewas carefully quenched with aqueous NH₄Cl solution, and extracted twicewith EtOAc. The combined organic layers were washed with saturatedNaHCO₃, brine, dried over Na₂SO₄, filtered, concentrated, and theresidue was purified by column chromatography on silica gel(EtOAc/Hexane: 40/60) to give 37a (106 mg, 71%) as yellow solid. ¹H NMR(500 MHz, CDCl₃) δ 3.01 (s, 2H), 3.19-3.33 (m, 2H), 4.43 (t, J=7.9 Hz,1H), 6.50 (d, J=8.3 Hz, 1H), 7.01-7.41 (m, 9H), 7.91 (s, 1H); MS calcdfor C₂₂H₂₀BrFN₂S (MH⁺) 456.06, found 456.1.

General Procedure for the Preparation of Sulfoxides and Sulphones

To a solution of the corresponding thioether in dry CH₂Cl₂ (5.0 ml) wasadded m-chloroperoxybenzoic acid (2-3 equiv.) at 0° C. under argon. Thereaction mixture was then stirred at 0° C. was allowed to warm to rtovernight. The reaction mixture was diluted with CH₂Cl₂ and was washedwith saturated NaHCO₃, brine, dried over Na₂SO₄, filtered, concentrated,and the residue was purified by column chromatography on silica gel.

38a (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R₃=3-methyl-1,2,4-oxadiazol-5-yl, m=1): obtained from 22a (70 mg, 0.18mmol) as a white solid (41 mg, 56. %). ¹H NMR (500 MHz, CDCl₃) δ 2.45(s, 3H), 3.53-3.67 (m, 2H), 4.18 and 4.32 (AB q, J=14.3 Hz, 2H), 5.01(dd, J=10.0 Hz, 5.5 Hz, 1H), 6.63 (d, J=8.9 Hz, 1H), 6.99 (d, J=8.9 Hz,1H), 7.05 (dd, J=8.4 Hz, 2.8 Hz, 1H), 7.24-7.29 (m, 3H), 7.42 (d, J=7.9Hz, 1H); MS calcd for C₁₈H₁₈Cl₂N₃O₂S (MH⁺) 410.05, found 409.9.

38b (W₁=5-Cl, R₁=2-chlorophenyl, R₂=NH₂,R₃=3-(4-fluorophenyl)-1,2,4-oxadiazol-5-yl, m=0): obtained from 14a.LC-MS: calcd. For C₂₂H₁₆Cl₂FN₃O₂S: 475.03; found: 497.9 [M+Na]⁺.

38c (W₁=5-Cl, R₁=2-chlorophenyl, R₃═NH₂,R₃=[(2,2-diphenylethyl)amino]carbonyl, m=0): obtained from 13i. LC-MS:calcd. For C₂₉H₂₆Cl₂N₂O₂S: 536.1; found: 558.9 [M+Na]⁺.

38d and 39d (W₁=5-Cl, R₁=2-chlorophenyl, R₂=H,R₃=[(cyclohexymethyl)amino]carbonyl, m=0) were obtained from 13aj. 38d:LC-MS: calcd. For C₂₂H₂₅Cl₂NO₂S: 437.1; found: 437.8 [M+H]⁺. 39d:C₇₇H₂₅Cl₂NO₃S: 453.1, found: 453.8 [M+H]⁺.

38e (W₁=5-Br, R₁=2-fluorophenyl, R₂=NH₂, R₃=(hydroxyamino)carbonyl,m=0): isolated as a side product in the synthesis of 2 ml (79 mg, 33%)as white solid: ¹H NMR (500 MHz, CDCl₃) δ 3.38-3.63 (m, 4sH), 4.71 (t,J=8.00 Hz, 1H), 6.52 (d, J=8.5 Hz, 1H), 6.97 (t, J=9.2 Hz, 1H),7.04-7.13 (m, 3H), 7.23-7.27 (m, 2H); MS calcd. for C₁₆H₂₇BrFN₂O₃S (MH⁺)415.01, found: 414.9.

Synthesis of 39

To a solution of 16o (0.035 g, 0.071 mmol) in dry CH₂Cl₂ (5 mL), DIPEA(0.049 mL, 0.28 mmol) was added and stirred at 0° C. A solution of MsCl(0.016 mL, 0.21 mmol) in dry CH₂Cl₂ (2 mL) was added dropwise slowlyover 30 min and the reaction was stirred for additional 3 hrs. Thereaction was diluted with CH₂Cl₂ (20 mL) and washed with a saturatedsolution of NaHCO₃ (2x10 mL), water (10 mL) and brine (20 mL). Theorganic layer was dried over sodium sulfate, filtered and concentratedunder vacuum. The crude product was purified by flash chromatography ona silica gel column using a mixture of petroleum ether and ethyl acetateas eluent to give 39 as a semi-transparent solid (0.032 g, 80% yield):LC/MS calcd for C₂₆H₃₅ClN₂O₆S₂: 570 [M−C₁₂H_(16N)O₄S], found: 300.

Synthesis of 40

Using similar procedures as for 15l and L8f, 40 was obtained from 10akand 15la (72% yield): LC/MS calcd for C₂₇H₃₇ClN₂O₄S₂: 552 [M+Na⁺],found: 575.

Synthesis of 41a and 41b

41a (X=Cl)

Using a similar procedure as for 15l, 41a was obtained from 10ak (84%yield): LC/MS calcd for C₂₉H₃₉ClN₂O₄S: 546 [M+Na⁺], found: 569.

41b (X=F)

Using a similar procedure as for 15l, 41b was obtained from 10ak (85%yield): LC-MS calcd for C₂₉H₃₉FN₂O₄S: 530 [M+Na⁺], found: 553.

EXAMPLE 15 Representative Compounds

The compounds listed in the following Table 1 were made by theprocedures disclosed in Examples 1-14 above. TABLE 1 RepresentativeCompounds Compound Structure & Number

11a

11aa

11ab

11ac

11ad

11ae

11af

11ag

11ah

11ai

11aj

11ak

11al

11am

11an

11b

11ba

11c

11d

11e

11f

11g

11h

11i

11j

11k

11l

11m

11n

11o

11p

11r

11s

11t

11u

11v

11w

11x

11y

12p

12r

12s

12

12x

13a

13ag

13ai

13aj

13b

13c

13d

13e

13f

13g

13h

13i

13j

13k

13l

13m

13n

13o

13p

13q

13r

13s

13t

13u

13v

14a

14ac

14ad

14b

14d

14g

14h

14i

14j

14k

14l

14la

14m

14ma

14o

14p

14q

14t

14w

14wa

14y

14z

15a

15b

15c

15d

15e

15ea

15f

15g

15h

15i

15j

15k

15l

16a

16b

16c

16d

16e

16f

16g

16h

16i

16j

16k

16l

16m

16n

16o

16p

16q

16r

16s

16t

16u

16v

16w

16x

16y

16z

16aa

16ab

16ac

16ad

16ae

17a

17b

17c

17d

17e

17f

17g

17h

17i

17j

17k

18a

18b

18c

18d

18e

18f

18g

20a

20b

20c

20d

21a

21b

21c

21d

21e

21f

21g

21h

21i

21j

22a

22b

22c

22d

25a

25b

25c

25d

26a

27a

27b

28a

28b

29a

29b

30a

30b

31a

34a

34b

34c

34d

34e

35f

36a

36b

36c

37a

38a

38b

38c

38d

38e

39

40

41a

41b

EXAMPLE 16 Mitochondrial Calcium/Sodium Antiporter Inhibitor PromotesEnhanced Insulin Secretion by Insulin-Secreting Cells COMPARATIVEEXAMPLE

INS-1 rat insulinoma cells were provided by Prof. Claes Wollheim,University Medical Centre, Geneva, Switzerland, and cultured at 37° C.in a humidified 5% CO₂ environment in RPMI cell culture media (GibcoBRL, Gaithersburg, Md.) supplemented with 10% fetal bovine serum (IrvineScientific, Irvine, Calif.), 2 mM L-glutamine, 100 U/ml penicillin, 100μg/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate and 50 μMβ-mercaptoethanol (all reagents Sigma, St. Louis, Mo., unless otherwisenoted).

INS-1 cells were seeded into 24-well plates containing RPMI mediasupplemented as described at 0.5×10⁶ cells/well and cultured at 37° C.,5% CO₂ for 2 days. Cells at or near confluence (0.7×10⁶ cells/well) wererinsed with glucose-free KRH buffer (134 mM NaCl, 4.7 mM KCl, 1.2 mMKH₂PO4, 1.2 mM MgSO₄, 1.0 mM CaCl₂, 10 mM HEPES-pH 7.4, 25 mM NaHCO₃,0.5% BSA), then incubated in the same buffer for 1 hr at 37° C. in ahumidified 5% CO₂/95% air atmosphere. Fresh KRH buffer was then added,either without added glucose (basal) or containing 8 mM glucose, in theabsence or presence of CPG37157—a known potent inhibitor of MCA(7-Chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2 (3H)-one)(Tocris Cookson, Inc., Ballwin, Mo.); see, e.g., Cox et al., 1993 TrendsPharmacol. Sci. 14:408; Maechler et al. 1997 EMBO J. 16:3833; Cox et al1993 J. Cardiovasc. Pharmacol. 21:595; White et al., 1997 J. Physiol.498:31; Baron et al., 1997 Eur. J. Pharmacol. 340:295; for relatedcompounds see, e.g., Chiesi et al., 1988 Biochem. Pharmacol. 37:4399).After an additional incubation for 15, 30 or 60 minutes at 37° C., 5%CO₂, the culture supernatants were collected. Insulin concentrations inthe supernatants were measured and normalized to cell number using aninsulin-specific radioimmunoassay kit (ICN Biochemicals, Irvine, Calif.)according to the manufacturer's instructions. The results are shown inFIG. 1, which illustrates enhanced glucose stimulated insulin secretionby INS-1 cells when exposed to CPG37157. FIG. 2 shows results that wereobtained when rat pancreatic islet cells were cultured under similarconditions in the presence of “basal” (5 mM) or supraphysiological (8mM) glucose, and in the absence or presence of various concentrations ofCPG37157.

EXAMPLE 17 Inhibition of Mitochondrial Calcium/Sodium AntiporterActivity

INS-1 rat insulinoma cells (see Example 16) were harvested bytrypsinization, washed and resuspended at 10×10⁶ cells/ml in assaybuffer (250 mM sucrose, 10 mM HEPES, 2.5 mM K₂HPO₄, 5 mM succinate, pH7.4) containing 0.007% digitonin, 60 μM CaCl₂ and 0.05 μM calcium green5N (Molecular Probes, Inc., Eugene, Oreg.). After a five-minute calciumloading incubation, ruthenium red (1 μM; Sigma, St. Louis, Mo.) wasadded to block further calcium uptake by mitochondria. Cell suspensionswere dispensed into 96-well plates (100 μl per well, 1×10⁶ cells perwell) and candidate agents (i.e., candidate mitochondrial calcium/sodiumantiporter inhibitors) were added to some sets of triplicate wells atconcentrations of 1, 10 or 100 μM, while other sets of wells providedappropriate control conditions (e.g., buffer and vehicle controls).Baseline fluorescence measurements were made using a multiwell platefluorimeter (F-MAX™, Molecular Devices Corp., Sunnyvale, Calif.; orPolarStar™, BMG Labtechnologies, Inc., Durham, N.C.) according to themanufacturer's instructions. Calcium efflux from mitochondria was theninduced by adding NaCl to all wells to achieve a final concentration of20 mM, and the rate of change in fluorescence in each well was monitoredwas monitored for two minutes and quantified using software includedwith the plate reader. Wells exhibiting significantly decreased changesin fluorescence over time relative to control wells indicated thepresence of agents that were candidate MCA inhibitors, and IC₅₀ valueswere calculated for these compounds. Preferred compounds of thisinvention have an IC₅₀ value of less than 10 μM, and more preferablyless than 1 μM. To that end, preferred compounds are listed in Table 2,while more preferred compounds are listed in Table 3. TABLE 2 IC₅₀ ≦ 10μM Compound Number 11aa 11am 11an 11ba 11c 11d 11f 11g 11h 11i 11k 11l11m 11r 11s 11t 11u 11w 11x 13a 13ag 13ai 13b 13c 13d 13e 13f 13g 13h13i 13j 13k 13l 13m 13n 13o 13p 13q 13r 13s 13t 13u 13v 14a 14b 14d 14g14h 14i 14k 14l 14m 14ma 14t 15c 15d 15e 15f 15i 15k 16a 16b 16c 16d 16e16f 16g 16h 16i 16j 16k 16l 16m 16p 16v 16y 17a 17b 17d 17e 17f 18b 18c18d 18e 20a 20b 20c 20d 21a 21d 21e 21f 21g 21h 22a 22b 22c 22d 25a 25b25c 25d 26a 26b 27b 28a 34c 35f 37a 38a 38c 39a 41a

TABLE 3 IC₅₀ ≦ 1 μM Compound Number 11aa 11ak 11ba 11d 11g 11h 11s 13b13c 13d 13f 13g 13h 13i 13j 13k 13l 13m 13n 13o 13p 13r 13s 13u 13v 14b14g 14h 14k 15c 15d 15e 15f 15i 15k 16a 16b 16d 16e 16f 16g 16h 16i 16j16k 16l 16m 16n 16o 16q 16r 16s 16t 16u 16w 16x 16z 16aa 16ab 16ac 16ad16ae 17b 17f 17g 17h 17i 17j 17k 18b 18c 18e 18f 18g 20b 20c 20d 21d 21e21f 21h 22a 22b 22d 25b 25d 26b 27b 38c 40 41b

EXAMPLE 18 Stimulation of Glucose-Stimulated Insulin Secretion

Pancreatic islets of Langerhans were isolated from adult maleSprague-Dawley rats using a standard collagenase infusion and digestionprocedure as described in Example 16. Islets were cultured at 37° C. for1-2 days in CMRL-1066 medium supplemented with 5.5 mM glucose, 10% fetalbovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, in ahumidified atmosphere containing 5% CO₂. Islets were manually picked andwashed in Krebs Ringer Bicarbonate buffer (KRB: 134 mM NaCl, 4.7 mM KCl,1.2 mM KH₂PO₄, 1.2 mM MgSO₄, 1 mM CaCl₂, 10 mM NaHCO₃, pH 7.4) inpreparation for measurement of glucose-stimulated insulin secretion(GSIS). Aliquots of washed islets were preincubated in oxygenated KRBsupplemented with 16 mM HEPES, 0.01% fetal bovine serum and 5.5 mMglucose for 60 min at 37° C. Compounds to be tested (e.g., candidatemitochondrial calcium/sodium antiporter inhibitors) were added atvarious concentrations for 10 minutes, after which additional glucosewas added to different islet cultures to achieve a final glucoseconcentration of 5.5, 8, 11 or 20 mM, and incubations were allowed toproceed an additional 20 min. Cell-conditioned media samples were thencollected by centrifugation and their insulin content was determinedusing enzyme-linked immunosorbent assay (ELISA) kits (CrystalChem orALPCO rat insulin ELISA) according to the kit supplier's instructions.Following treatment with a preferred compound, the concentration ofinsulin detected in the islet-conditioned medium was at least 1.5 timesthe insulin concentration detected in the medium conditioned by isletsthat were exposed to 8 mM glucose.

At a concentration of 1 μM, CGP37157 stimulated islet GSIS by 222 (48) %relative to GSIS detected with 8 mM glucose. Preferred compounds of thisinvention stimulate GSIS by 150% or more at a concentration of 1 μLM,and more preferably by 200% or more at a concentration of 1 μM. To thatend, preferred compounds are listed in Table 4, while more preferredcompounds are listed in Table 5. TABLE 4 Compound Number 11d 11g 11h 11k11t 11x 13t 14a 14b 16d 16d 16e 16f 16i 16k 17a 20a 20b 20c 21e 22a 25b25c 25d 27b 28b

TABLE 5 Compound Number 20a 11t 25c 25d 11g 11d 25b 11h 14a 11k 20b 20c13t 16d 16e 16k 17a

EXAMPLE 19 Effect of Oral Administration of a Representative Compound onGlucose Tolerance in Mutant DB/DB Mice

This Example illustrates the effects of a representative compound onglucose tolerance in an established animal model of type R diabetesmellitus, the db/db mutant mouse. As a brief background, the recessivedb mutation has been localized to murine chromosome 4, and homozygousrecessive (db/db) individuals are characterized by, inter alia, obesity,hyperphagia, transient increases in plasma insulin concentrations,hyperglycemia, abnormal immune and renal functions, diabetic neuropathyand myocardial disease (see, e.g., Hummel et al., 1966 Science 153:1127;Herberg et al., 1977 Metabolism 26:59; Leiter et al., 1981 Metabolism30:554; Guenet et al., 1982 Mouse News Letter 67:30; Guenet et al., 1984Mouse News Letter 70:95; Bray et al., 1971 Physiol. Rev. 51:598; Baileyet al., 1989 J. Endocrinol. 123:19-24; Bray et al., 1979 Physiol. Rev.59:719; Sima et al., 1979 Lab. Invest. 40:627; Sima et al., 1978 ActaNeuropathol. 41:85; Giacomelli et al., 1979 Lab. Invest. 40:460).

16o (dissolved in vehicle: 10% (v/v) EtOH, 10% (v/v) polyethyleneglycol-400, 30% (v/v) propylene glycol, 50% (v/v) H₂O) was administeredorally at a dosage of 100 mg/kg body weight to eight-week old mutantC57BLKs-db/db mice (Harlan Bioproducts for Science, Inc., Indianapolis,Ind.). After one hour, a bolus of glucose dissolved in sterile normalsaline solution (1 gm glucose/kg body weight) was injectedintraperitoneally (time 0, FIG. 3), and blood samples were collected at15, 30, and 90 minutes following the glucose load. Glucoseconcentrations in each blood sample were determined using OneTouch®glucose test strips (LifeScan, Inc., Milpitas, Calif.) according to themanufacturer's instructions. Control animals received theEtOH/PEG/propylene glycol/H₂O vehicle only, containing no 16o.

The results are presented in FIG. 3, which shows that relative to db/dbmice that received only the vehicle control, the db/db animals thatreceived 16o exhibited lower peak blood glucose levels after the glucosebolus, and returned to a lower baseline more rapidly (*p<0.05).

From the foregoing, it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1. A compound having the structure:

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof,wherein: X is —S(O)_(q)—, —O—, —N(R)— or —C(R)(R′)—; m is 0 or 1; n is0, 1 or 2; q is 0, 1 or 2; W₁ and W₂ each represent an optionalsubstituent, wherein W₁ and W₂ are the same or different andindependently halogen, nitro, or lower alkyl; R and R′ are the same ordifferent and independently alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl or substituted arylalkyl, or R and R′ taken togetherwith the carbon atom to which they are bonded form a carbocycle,substituted carbocycle, heterocycle or substituted heterocycle; R₁ isalkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl or substituted heteroaryl; R₂ is hydrogen, nitro,—OR_(2a), —C(═O)NR_(2b)R_(2c), —CH₂NR_(2b)R_(2c), —CH₂OR_(2a),—NR_(2b)R_(2c), —NHC(═O)R_(2a), —NHC(═O)NR_(2b)R_(2c) or—NHC(═NH)NR_(2b)R_(2c); R_(2a) is hydrogen, alkyl, substituted alkyl,arylalkyl, or substituted arylalkyl; R_(2b) and R_(2c) are the same ordifferent and independently hydrogen, alkyl, substituted alkyl, —SO₂R₄,—C(═NH)NH₂ or —C(═O)R_(2d) where R_(2d) is amino, alkyl, substitutedalkyl, aryl or substituted aryl; R₃ is hydroxy, alkyl, substitutedalkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl,heterocycle, substituted heterocycle, heterocyclealkyl, substitutedheterocyclealkyl, —C(═O)N(R_(3a))(R_(3b)), —NHC(═O)N(R_(3a))(R_(3b)),—NHC(═S)N(R_(3a))(R_(3b)), —C(═O)OR_(3c), —C(═O)R_(3c), —NHC(═O)R_(3d)or —NHSO₂R_(3d); R_(3a) and R_(3b) are the same or different andindependently hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl or substituted arylalkyl, or R_(3a) and R_(3b) takentogether with the nitrogen atom to which they are attached form aheterocycle or substituted heterocycle; R_(c) is hydrogen, alkyl,substituted alkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl orsubstituted heteroarylalkyl; R_(3d) is alkyl, substituted alkyl, aryl,substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl;and R₄ is, at each occurrence, the same or different and independentlyhydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl orsubstituted arylalkyl.
 2. The compound of claim 1 wherein X is—S(O)_(q)—.
 3. The compound of claim 1 wherein X is —O—.
 4. The compoundof claim 1 wherein X is —N(R)—.
 5. The compound of claim 1 wherein X is—C(R)(R′)—.
 6. The compound of claim 1 wherein R₂ is hydrogen.
 7. Thecompound of claim 1 wherein R₂ is —OR_(2a).
 8. The compound of claim 1wherein R₂ is —C(═O)NR_(2b)R_(2c).
 9. The compound of claim 1 wherein R₂is —CH₂NR_(2b)R_(2c).
 10. The compound of claim 1 wherein R₂ is—CH₂OR_(2a).
 11. The compound of claim 1 wherein R₂ is —NR_(2b)R_(2c).12. The compound of claim 11 wherein R_(2b) and R_(2c) are hydrogen. 13.The compound of claim 1 wherein R₂ is —NHC(═O)R_(2a).
 14. The compoundof claim 1 wherein R₂ is —NHC(═O)NR_(2b)R_(2c).
 15. The compound ofclaim 1 wherein R₂ is —NHC(═NH)NR_(2b)R_(2c).
 16. The compound of claim1 wherein R₁ is aryl or substituted aryl.
 17. The compound of claim 16wherein W₁ is present at the 5-position and W₂ is not present.
 18. Thecompound of claim 17 wherein W₁ is halogen.
 19. The compound of claim 18wherein W₁ is chloro.
 20. The compound of claim 18 wherein X isS(O)_(q), q is 0, R₂ is —NR_(2a)R_(2c), m is 0 and n is
 1. 21. Thecompound of claim 18 wherein X is S(O)_(q), q is 0, R₂ is—NR_(2a)R_(2c), m is 0 and n is
 2. 22. The compound of claim 20 or 21wherein R₃ is —C(═O)N(R_(3a))(R_(3b)).
 23. The compound of claim 20 or21 wherein R₃ is —C(═O)OR_(3c).
 24. The compound of claim 20 or 21wherein R₃ is —C(═O)R_(3c).
 25. The compound of claim 20 or 21 whereinR₃ heterocycle or substituted heterocycle.
 26. The compound of claim 20or 21 wherein R₃ is NHC(═O)N(R_(3a))(R_(3b)).
 27. The compound of claim20 or 21 wherein R₃—NHC(═S)N(R_(3a))(R_(3b)).
 28. The compound of claim20 or 21 wherein R₃ is —NHC(═O)R_(3d).
 29. The compound of claim 20 or21 wherein R₃ is —NHSO₂R_(3d).
 30. The compound of claim 1 wherein R₁ isalkyl or substituted alkyl.
 31. The compound of claim 30 wherein W₁ ispresent at the 5-position and W₂ is not present.
 32. The compound ofclaim 31 wherein W₁ is halogen.
 33. The compound of claim 32 wherein W₁is chloro.
 34. The compound of claim 32 wherein X is S(O)_(q), q is 0,R₂ is —NR_(2a)R_(2c), m is 0 and n is
 1. 35. The compound of claim 32wherein X is S(O)_(q), q is 0, R₂ is —NR_(2a)R_(2c), m is 0 and n is 2.36. The compound of claim 34 or 35 wherein R₃ is—C(═O)N(R_(3a))(R_(3b)).
 37. The compound of claim 34 or 35 wherein R₃is —C(═O)OR_(3c).
 38. The compound of claim 34 or 35 wherein R₃ is—C(═O)R_(3c).
 39. The compound of claim 34 or 35 wherein R₃ heterocycleor substituted heterocycle.
 40. The compound of claim 34 or 35 whereinR₃ is NHC(═O)N(R_(3a))(R_(3b)).
 41. The compound of claim 34 or 35wherein R₃—NHC(═S)N(R_(3a))(R_(3b)).
 42. The compound of claim 34 or 35wherein R₃ is —NHC(═O)R_(3d).
 43. The compound of claim 34 or 35 whereinR₃ is —NHSO₂R_(3d).
 44. A method for treating diabetes mellitus,comprising administering, to a subject having or suspected of being atrisk for having diabetes mellitus, a therapeutically effective amount ofa pharmaceutical composition comprising a pharmaceutically acceptablecarrier and a compound of claim
 1. 45. The method of claim 44 whereinthe diabetes mellitus is type 2 diabetes mellitus.
 46. The method ofclaim 44 wherein the diabetes mellitus is maturity onset diabetes of theyoung.
 47. A method for enhancing insulin secretion, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a compound of claim
 1. 48. A method forinhibiting a mitochondrial calcium/sodium antiporter, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a compound of claim
 1. 49. The method of any oneof claims 44, 47 or 48, further comprising administration to the subjectone or more agents that lower circulating glucose concentration.